Real-time PCR quantification and histopathological findings of Mycoplasma hyopneumoniae infection in the lungs of pigs slaughtered in São Luís, Maranhão, Brazil / Quantificação por PCR em tempo real e achados histopatológicos da infecção por Mycoplasma hyopneumoniae em pulmão de suínos abatidos na cidade de São Luís, Maranhão, Brasil

ABSTRACT: Porcine enzootic pneumonia (PES), mainly caused by the bacteria Mycoplasma hyopneumoniae, is the main cause of respiratory problems in pigs. Infection by M. hyopneumoniae leads to production losses and the predisposition of affected animals to secondary infections, which may result in the condemnation of carcasses and organs due to lung lesions at the time of slaughter. The objective of the research was to evaluate the infection by M. hyopneumoniae in pigs submitted to slaughter in São Luís Island/MA, using molecular and histopathological diagnostic methods. One hundred fifty lung samples were collected from inspected (n=65) and non-inspected (n=85) slaughter pigs on São Luís Island, Maranhão, from July 2019 to August 2021. Of the 150 DNA samples collected, 121 showed an amplified product for Cyt B in the PCR assay. Thus, 121 samples were submitted to qPCR of M. hyopneumoniae, of which 44 (36.36%) showed positive results. The mean amount of bacterial load ranged from 1.20 × 101 to 7.20 × 104, with a mean of 1.73 × 104 copies. Of the reagent samples, 81.81% (36 samples) were obtained from non-inspected slaughter, while 18.18% (8 samples) were obtained from slaughterhouses. In the histopathological analysis, 44 positive qPCR samples were evaluated, of which 28 (63.63%) presented results compatible with the main inflammatory process associated with the presence of M. hyopneumoniae, that is, bronchial-associated lymphoid tissue hyperplasia (BALT). Three samples that showed the highest bacterial load (qPCR: 5.63 × 10³, 2.19 × 104 and 7.23 × 104) showed more evident lesions in this study. The microscopic findings associated with the quantifications indicated a relationship between the amount of bacterial load and the presence of microscopic lesions; higher bacterial load in lung tissue is associated with increased histopathologic staining for BALT hyperplasia. In conclusion, the results point to the circulation of the etiological agent in the sampled animals and the need for preventive measures on pig farms in Maranhão with the involvement of producers, sanitary defense and inspection agencies.

RESUMO: A pneumonia enzoótica suína (PES), causada principalmente pela bactéria Mycoplasma hyopneumoniae, é a principal causa de problemas respiratórios em suínos. A infecção por M. hyopneumoniae leva a perdas produtivas e a predisposição dos animais acometidos a infecções secundárias, o que pode resultar em condenação de carcaças e órgãos por lesões pulmonares no momento do abate. O objetivo da pesquisa foi avaliar a infecção por M. hyopneumoniae em suínos submetidos ao abate na Ilha de São Luís, por meio de métodos diagnósticos moleculares e histopatológicos. Para isso, foram coletadas 150 amostras de pulmão de suínos de abate inspecionado (n=65) e não inspecionado (n=85) na Ilha de São Luís/Maranhão, no período de julho de 2019 a agosto de 2021. Das 150 amostras de DNA coletadas, 121 apresentaram produto amplificado para Cyt B no ensaio de PCR. Assim, 121 amostras foram submetidas à qPCR de M. hyopneumoniae, das quais 44 (36,36%) apresentaram resultados positivos. A quantidade média de carga bacteriana variou de 1,20 × 101 a 7,20 × 104, com média de 1,73 × 104 cópias. Das amostras reagentes, 81,81% (36 amostras) foram obtidas de abate não inspecionado, enquanto 18,18% (8 amostras) foram obtidas em abatedouro. Na análise histopatológica, foram avaliadas 44 amostras positivas para qPCR, das quais 28 (63,63%) apresentaram resultados compatíveis com o principal processo inflamatório associado à presença de M. hyopneumoniae, ou seja, hiperplasia do tecido linfóide associado ao brônquio (BALT). Três amostras que apresentaram maior carga bacteriana (qPCR: 5,63 × 10³, 2,19 × 104 e 7,23 × 104) foram mais evidentes neste estudo. Os achados microscópicos associados às quantificações indicaram uma relação entre a quantidade de carga bacteriana e a presença de lesão microscópica; a maior carga bacteriana no tecido pulmonar está associada a maior alteração histopatológica para hiperplasia BALT. Em conclusão, os resultados obtidos sinalizam para a circulação do agente etiológico nos animais amostrados e a necessidade de medidas preventivas nas criações de suínos do estado do Maranhão com envolvimento dos produtores, órgãos de defesa sanitária e inspeção.

Deeper exploration of inflammatory cell populations in milk to monitor udder health in dairy cows

The present study explored the predictive values of milk leukocyte differentials (MLD) as a basis for improving the diagnosis of intramammary infections (IMIs) and subclinical mastitis. Quarter milk samples were collected for bacteriological analysis, quarter somatic cell count (qSCC), and MLD. The MLD were assessed using the cytospin technique, direct microscopic smears, and flow cytometry. The predictive values of each single leukocyte population and useful potential indices that could better reflect immune complexity were also calculated. Changes in the percentage of any leukocyte alone failed to substantially improve the predictive value of qSCC in diagnosing IMIs. Although certain parameters increased the area under the receiver operating characteristic curve (ROC curve) as a result of increased specificity values, a slight reduction in sensitivity was observed. The so-called CD8 complex was a unique parameter which improved both the sensitivity (78.79 %) and the specificity (80.77 %) in IMI diagnosis, resulting in the highest area under the ROC curve (0.87). To diagnose subclinical mastitis, the percentage of macrophages and the sum of the percentage PMNLs and T CD8+ cells divided by the percentage of macrophages showed the highest predictive values (sensitivity = 79.63, specificity = 73.68, and area under the ROC curve = 0.83) in the differentiation of the inflammatory condition status of cows. In conclusion, this study provides further insights into using T CD8+ lymphocytes in diagnosing bovine IMIs, combined with PMNLs and macrophages. The antidromic trend of macrophages vs. PMNLs and T CD8+ lymphocytes due to the increasing qSCCs was crucial to differentiating quarters under both inflammatory and non-inflammatory conditions.

Clinical, histopathological, and hematological changes due to isoimmune thrombocytopenic purpura in piglets / Alterações clínicas, histopatológicas e hematológicas decorrentes de púrpura trombocitopenica isoimune em leitões

ABSTRACT: Isoimmune thrombocytopenic purpura (ITP) is an immune-mediated disease that causes severe hemorrhagic lesions and high mortality in piglets. The disease can occur early in newborn piglets (EITP) or late in 2- to 3-week old piglets (LITP). In this study, we analysed the clinical, pathological, and hematological aspects of 391 ITP cases (312 with EITP and 79 with LITP). In LIPT cases, morbidity and mortality rates were higher, with rates of 60% (morbidity) and 53% (mortality). The main clinicopathological findings in ITP cases were different patterns of hemorrhages organs and tissues. In EITP, clinical signs were characterized by extensive subcutaneous hemorrhages and death occurred within a few days; however, in LITP, often sudden death occurred. In macroscopic analysis, hemorrhagic diathesis was observed in all affected animals. In EITP, the most severe hemorrhagic lesions were integumentary, mainly in the dermis and epidermis. In LITP, visceral lesions were predominant, mainly in the epicardium and intestines. Microscopic bone marrow analysis revealed mild cellular hyperplasia in EITP and bone marrow aplasia in LITP. hematological analyses revealed leucopenia, thrombocytopenia, and anemia in all ITP-affected animals. However, fostering by a different sow was only efficient in controlling EITP and had little effect in LITP-symptomatic piglets, due to more severe lesions. Further studies on the etiopathogenesis of LITP are required to improve our understanding of this disease form.

RESUMO: Púrpura trombocitopênica isoimune (PTI) é uma doença imunomediada que causa lesões hemorrágicas graves e alta mortalidade em leitões, que pode se apresentar através de uma forma precoce em leitões neonatos (PTIP) ou uma forma tardia em leitões com duas a três semanas de idade (PTIT). Neste trabalho analisamos aspectos clínicos, hematológicos e histopatológicos de 391 casos de PTI, sendo 312 de PTIP e 79 de PTIT. Observou-se maiores morbidade (60%) e mortalidade (53%) na PTIT. Os principais achados clínico-patológicos observado na PTI são hemorragias em diferentes graus de intensidade e nos diferentes órgãos e tecidos. Na PTIP observou-se predominantemente hemorragias subcutâneas extensas e morte em alguns dias, já na PTIT, observou-se além de grave hemorragia, morte súbita. Na análise macroscópica, observou-se diátese hemorrágica em todos os animais afetados. Na PTIP as lesões hemorrágicas mais graves foram tegumentares, principalmente em derme e epiderme, enquanto, na forma tardia, observou-se lesões predominantemente viscerais, em epicárdico e intestino. A análise microscópica de medula óssea revelou discreta hiperplasia celular na forma PTIP, enquanto, na PTIT observou-se aplasia medular. Na análise hematológica observou-se leucopenia, trombocitopenia e anemia em todos os animais com PTI. Os achados clínicos, histopatológicos e hematológicos para PTIP e PTIT da doença permitiram o diagnóstico de PTI. Entretanto, a troca de mãe se mostrou eficiente apenas para controle PTIP, uma vez que, esta estratégia apresenta pouco resultado para leitões sintomáticos com a PTIT, devido lesões mais severas. Estudos sobre a etiopatogênese da PTIT ainda são necessários para melhor entendimento desta forma da doença.

Aspects of skin morphology and morphometry of the giant anteater (Myrmecophaga tridactyla, LINNAEUS, 1758) / spectos da morfologia e morfometria da pele do tamanduá bandeira (Myrmecophaga tridactyla, LINNAEUS, 1758)

The aim of the present study is to describe the skin morphology and morphometry of the giant anteater (M. tridactyla), based on comparative analysis applied to skin segments from central metacarpal torus (palmar pad), dorsal thorax, ventral cervical, ventral abdomen, medial carpal and nasal regions. In order to do so, eight adult specimens of M. tridactyla were used for macroscopic studies and four for microscopic assessments. Microscopy was used to assess fur general features that were macroscopically assessed through visual analysis. Fragments (2.0cm²) were collected from the selected regions for microscopic studies. Samples were fixed on McDowell solution, processed through routine histology techniques and subjected to semi-serial cuts (5 µm). The cuts were stained in HE, Alcian blue and periodic acid Schiff. General morphology of different skin layers was described, as well as their architecture and composition; mesoscopy of the epidermis, dermis and stratum corneum was also carried out. There was difference in skin morphometry between males and females, and between different skin regions in the same animal, based on the statistical evaluation of the recorded values. All epidermis layers were assessed for the selected regions. Dermis encompassed surface and deep layers; it presented sweat and sebaceous glands, as well as hair follicles. Findings also allowed reporting that epidermis components are easily identified given its thickness, and the large amount of sweat glands in it ­ it contrasts its physiological features.(AU)

O objetivo deste estudo foi descrever a morfologia e morfometria da pele do Tamanduá-bandeira (M. tridactyla), mediante a análise comparativa de segmentos cutâneos das regiões central do toro metacarpal (coxim palmar), dorsal do tórax, cervical ventral, ventral do abdome, medial do carpo e nasal. Para tanto, foram utilizados oito exemplares adultos de M. tri-dactyla para o estudo macroscópico, e quatro destes para o estudo microscópico. Macroscopicamente estudou-se, por meio de análise visual, as características gerais do pelo e pelagem. Para o estudo microscópico, foram coletados fragmentos de 2,0 cm2 das áreas selecionadas. As amostras foram fixadas em solução de McDowell, processadas pelas técnicas rotineiras de his-tologia e submetidas a cortes semi-seriados de 5 µm. Os cortes foram corados com HE, azul de Alcian e ácido periódico de Schiff. Descreveu-se a morfologia geral, a arquitetura e a composição das diferentes camadas da pele e ainda realizou-se a mesoscopia da epiderme, derme e estrato córneo. De acordo com a avaliação estatística dos valores obtidos, houve diferença na morfometria cutânea entre machos e fêmeas, e entre as diferentes regiões cutâneas de um mesmo animal. Nas regiões estu-dadas observou-se todas as camadas da epiderme. A derme compôs-se das camadas superficial e profunda, apresentado glân-dulas sudoríferas e sebáceas e folículos pilosos. Dentre os achados também pode-se relatar que os componentes da epiderme são facilmente identificáveis pela sua maior espessura, além da grande quantidade de glândulas sudoríferas presentes, o que contrasta com suas características fisiológicas.(AU)

Crescimentos de fungos produtores de micotoxinas em grãos de amendoim / Growths of mycotoxin-producing fungi on peanut grains

O amendoim (Arachis hypogaea L.) é uma oleaginosa com alto valor energético e nutricional, servindo de substrato ideal para o crescimento de fungos durante o processo de colheita, estocagem e processamento dos grãos. As micotoxinas são originadas a partir do metabolismo secundário de fungos toxigênicos; quando essas toxinas são ingeridas, podem causar efeitos tóxicos em humanos e animais. O objetivo deste trabalho foi avaliar a contaminação fúngica em amostras de grãos de amendoim disponíveis para o consumo humano. Foram analisadas amostras de amendoim torrado na vagem, amendoim cru (in natura) e amendoim torrado sem a vagem. A metodologia utilizada foi o método de plaqueamento direto em placas de Petri, contendo o meio cultura Sabouraud Dextrose Ágar (SDA). Porções de 40 grãos de amendoim de cada amostra foram utilizadas para o teste. Os resultados obtidos foram expressos em quantidade de grãos que apresentaram crescimento fúngico em porcentagem. Para identificação ao nível do género das colônias fúngicas isoladas, foi efetuada uma observação microscópica das suas estruturas morfológicas. Em todas as amostras estudadas ocorreram o crescimento de fungos potencialmente micotoxigênicos. Os grãos de amendoim cru (in natura) sem a vagem foram os mais contaminados por fungos como Aspergillus sp. (20%), Alternaria sp. (13,1%), Penicillium sp. (13,3%) e Fusarium sp. (23,3%) quando comparados com os grãos torrados. A presença desses fungos nas amostras de amendoim indica a possibilidade de desenvolvimento de micotoxinas potencialmente tóxicas para o humano. O processo de torrefação inibe o crescimento de determinadas espécies de fungos micotoxigênicos.(AU)

Peanut is an oilseed with high energy and nutritional value, serving as an ideal substrate for the growth of fungi during the process of harvesting, storing and processing the grains. Mycotoxins originate from the secondary metabolism of toxigenic fungi; when ingested, they can cause toxic effects in humans and animals. The objective of this work was to evaluate the fungal contamination in samples of peanut grains available for human consumption. The methodology used was the direct plating method in Petri dishes containing Sabouraud Dextrose Agar (SDA) culture. Portions of 40 peanut kernels from the samples were immersed in a 0.4% sodium hypochlorite solution for two minutes for surface disinfection, then rinsed once in distilled water. Incubation was carried out at room temperature for 7 days and the results were expressed as the amount of grains that showed fungal growth in percentage. To identify the colonies of isolated filamentous fungi at the genus level, a microscopic observation of their morphological structures was carried out, which consists of removing a sample from the edge of the colony and placing it in a drop of cotton blue (1g/L in lactic acid 88%), between slide and coverslip. Potentially toxigenic fungi were identified: Aspergillus sp., Rhizopus sp., Alternaria sp., Penicillium sp. and Fusarium sp. The presence of these fungi in peanut samples indicates the possibility of development of mycotoxins potentially toxic to humans. The roasting process may not be sufficient to destroy some mycotoxins.(AU)

Canine ringworm caused by Trichophyton mentagrophytes - detection by SYBR-Green real-time PCR

Background: Dermatophytes, fungi of universal distribution, invade semi or fully keratinized structures, such as skin, fur/ hair and nails. The various species of dermatophytes are classified into three genera anamorphic: Microsporum, Trichophyton and Epidermophyton. The genus Epidermophyton includes only E. floccosum, that rarely affects animals. The main species responsible for the disease in dogs and cats are Microsporum canis, M. gypseum and Trichophyton mentagrophytes, which were characterized through conventional mycological methodology (microscopic examination with KOH and culture). Molecular methodologies, such as real-time PCR, can contribute to a rapid laboratory diagnosis, helping clinicians to initiate an early antifungal treatment. This case report describes a case of canine dermatophytosis due to Trichophyton mentagrophytes detected from a clinical sample by SYBR-Green real-time PCR. Case: A 8-year-old dog, rescued from the street, was referred to a private veterinary clinic in the city of Canoas, RS, Brazil, presenting generalized lymphadenomegaly, crusted lesions all over the body, generalized alopecia, signs of excoriation and epistaxis. Initially, were administered prednisone [1 mg/kg every 48 h, BID] and cephalexin [30 mg/kg, BID]. Weekly baths with benzoyl peroxide were also given. The therapy was not clinically successful. Wood's Lamp Test was negative. As a differential diagnosis, PCR for detection of Leishmania was negative. Complete blood count and serum biochemical assay were also performed. For mycological diagnosis, hair specimen was clarified and examined microscopically using 10% potassium hydroxide (KOH) for the visualization of chains of arthroconidia (ectothrix invasion of hair). The infected hair was plated onto MycoselTM Agar, incubated at 28°C for 15 days. Microscopy of hyphae/ conidia and macroscopic colony characteristics (colors and texture) were conducted for the differentiation of the species within the genus Microsporum and Trichophyton. In addition, real-time PCR was applied for direct analysis of the fungal DNA obtained from the hair sample. Microscopic examination was negative. The dermatophyte present in the hair sample was confirmed as Trichophyton mentagrophytes by culture and qPCR (melting-point analysis). The patient was treated with systemic itraconazole [10 mg/ kg SID - 90 days]. Twice-weekly application of 2.5 % miconazole and 2% chlorhexidine shampoo until complete cure. Discussion: Dermatophytosis is often listed as self-limiting infection; however, animal dermatophytosis can spread between pets, as well as a zoonotic transmission to humans. The literature on dermatophytosis indicates that Microsporum canis is the predominant etiological agent, followed by M. gypseum. Trichophyon mentagrophytes that appear in a lower percentage of isolation. The culture of hair, even with specific medium containing chloramphenicol and cyclohexamide, may present contaminating fungi, not related to dermatophytosis, which can inhibit or override the growth of dermatophytes. The use of real-time PCR provided a faster and specific diagnosis of dermatophytosis when compared to the conventional mycological methodology for detection and identification of T. mentagrophytes, which takes around 10 to 15 days for culture. It is possible to use this technique as an alternative diagnosis for dermatophytes associated to clinical hair samples of dogs.

Gota úrica visceral em pato-do-mato (Cairina moschata) / Visceral uric gout in muscovy duck (Cairina moschata)

Background: Gout, a metabolic disease affecting multiple species, is frequent in birds which are uricotelic animals. It is primarily caused by hyperuricemia originating from birds' renal disorders with subsequent deposition of urate crystals into tissue. The location of the crystals determines the type of disease. Joint disease occurs with deposition inside and around joints, joint capsules, and tandine hems, while visceral disease occurs when the serous surface of visceral organs (mainly pericardium, liver, kidneys, air sacs, peritoneum, and spleen) are affected. This report describes a case of visceral uric gout in a Muscovy duck, which is rare. Case: An adult, male, Muscovy duck (Cairina moschata), kept under human care in a zoo, was referred to a veterinary clinic. The animal presented suddenly with prostration, ataxia, and anorexia, and died the following day. The body was sent to the Pathology Sector of Ritter dos Reis University Center (UniRitter) for a necropsy. During the necropsy, it was observed that white membranes, presenting a "chalk dust" appearance, were deposited under the serous layer of organs including the liver, kidneys, and pericardium. Following the necropsy and macroscopic evaluations, fragments of multiple organs were fixed in 10% formalin and processed routinely with hematoxylin and eosin (HE) staining for histopathological evaluation in an analysis laboratory in Porto Alegre city. A deposition of amorphous eosinophilic material was identified in the intestine, liver (serous), kidneys, lungs, and heart (epicardium) with findings of serositis, nephritis, and fibrinous epicarditis. The liver parenchyma and lungs had areas of congestion. There was both moderate and intense hepatocellular degeneration as well as degeneration in the renal tubular cells. These macro and microscopic changes were compatible with urate crystal deposition which represents visceral uric gout. Discussion: As opposed to free-living birds, birds in captivity have easy access to food and are sedentary. These factors, together with inadequate food management, make nutritional disorders the main predisposing cause of uric gout in these animals. Since it was not possible to determine the causative factors of this individual animal's disease due to the absence of examinations while alive, a nutritional origin of this animal's demise was considered. According to prior reports, an important cause of avian disease can be attributed directly or indirectly to kidney dysfunction. In addition to a deposition of amorphous material in the renal cortex, this Muscovy duck had areas of tubular cell degeneration and proliferation of fibrous connective tissue. However, it was not possible to establish a cause and effect relationship between the renal damage and the uric gout. Despite being common in birds under human care, the disease still is challenging for veterinarians, since the diagnosis is often late and treatment is often ineffective. The absence of documentation of this disease in the Muscovy duck contributes to the difficulty in establishing predisposing factors and a distinct disease etiology. This highlights the importance of the present report and the need for new studies. To the best of our knowledge, this is the first report of visceral uric gout in a Muscovy duck (Cairina moschata). This highlights the importance of necroscopic examinations and the impact they can have in the clinical arena, especially in wild animals.

Eficiência de três diluidores inespecíficos na refrigeração do sêmen de coelho (Oryctolagus cuniculus f. domesticus) / Efficiency of three unspecific diluents in refrigeration of rabbit (Oryctolagus cuniculus f. domesticus) semen / Eficiencia de tres diluyentes inespecíficos en la refrigeración de semen de conejo (Oryctolagus cuniculus f. domesticus)

Objetivou-se com este trabalho avaliar a viabilidade seminal de coelhos após refrigeração a 5°C utilizando três diluentes comerciais, indicados para refrigeração de sêmen de outras espécies monogástricas. Foram realizadas dez colheitas de sêmen de cada reprodutor. Os ejaculados de cada coelho eram misturados, obtendo-se um pool de sêmen. Em seguida, o pool heterospérmico foi dividido em três tratamentos, de acordo com o diluente utilizado: BotuDogⓇ (cães); BotuSemen® (equinos); e Beltsville Thawing Solution - BTS (suínos). As amostras foram avaliadas quanto aos parâmetros macroscópicos, microscópicos e funcionais em quatro momentos: sêmen fresco (0 h) e refrigerado (16, 24 e 48 h). Os valores médios de motilidade, vigor e defeitos totais do sêmen fresco foram de 86,0%, 3,05 e 11,5%, respectivamente. Após o início do resfriamento, observou-se comportamento linear decrescente para a motilidade e vigor espermático do sêmen em função do tempo, diferindo (P < 0,05) entre os tratamentos. Destaca-se que o tratamento diluído em BTS apresentou redução na manutenção dos parâmetros avaliados de forma mais pronunciada (P < 0.05) ao longo do tempo, apresentando os menores valores para motilidade e vigor espermático dentro de cada tempo avaliado: 16h [20,6 ± 7,28 e 0,75 (0,375 1,5)], 24h [12,0 ± 6,80 e 0 (0 - 1,0)] e 48h [3,0 ± 2,13 e 0 (0 - 0,125)]. Após 48 h de resfriamento, o BotuDogⓇ foi o que apresentou maior valor para motilidade espermática (71,0 ± 2,08) e, juntamente, com o BotuSemenⓇ apresentaram maior valor para vigor espermático (2,5). De acordo com os dados obtidos os diluentes BotuDogⓇ e BotuSemenⓇ mostraram-se eficientes em manter parâmetros espermáticos adequados, apresentando boa viabilidade espermática em até 48h pós refrigeração a 5°C. Foram obtidos, resultados inéditos quanto a parâmetros espermáticos do sêmen de coelho refrigerado que podem ser utilizados como valores de referência para o manejo reprodutivo e processamento do semen desses animais, visto a inexistênca dessas recomendações técnicas.

The objective of this work was to evaluate the seminal viability of rabbits after refrigeration at 5°C using three commercial diluents, indicated for refrigeration of semen from other monogastric species. Ten semen collections were performed from each breeder. The ejaculates of each rabbit were mixed, obtaining a pool of semen. Then, the heterospermic pool was divided into three treatments, according to the diluent used: BotuDog (dogs); BotuSemenⓇ (horses); and Beltsville Thawing Solution - BTS (pigs). The samples were evaluated for macroscopic, microscopic, and functional parameters in four moments: fresh (0 h) and refrigerated (16, 24, and 48 h) semen. The average values of motility, vigor, and total defects of fresh semen were 86.0%, 3.05 and 11.5%, respectively. After the beginning of cooling, a decreasing linear behavior was observed for motility and sperm vigor of semen as a function of time, differing (P < 0.05) between treatments. It is noteworthy that the treatment diluted in BTS® showed a more pronounced reduction in the maintenance of the evaluated parameters (P < 0.05) over time, with the lowest values for sperm motility and vigor within each evaluated time: 16h [20.6 +7.28 and 0.75 (0.375 - 1.5)], 24h [12.0± 6.80 and 0 (0 - 1.0)] and 48h [3.0±2.13 and 0 (0 - 0.125)]. After 48 h of cooling, BotuDog presented the highest value for sperm motility (71.0 ± 2.08) and, together with BotuSemenⓇ, presented the highest value for sperm vigor (2.5). According to the data obtained, BotuDog and BotuSemenⓇ diluents were efficient in maintaining adequate sperm parameters, showing good sperm viability in up to 48 hours after refrigeration at 5°C. Through this work, unpublished results were obtained regarding spermatic parameters of refrigerated rabbit semen that can be used as reference values for reproductive management and semen processing of these animals, given the lack of these technical recommendations.

El objetivo de este trabajo fue evaluar la viabilidad seminal de conejos después de refrigeración a 5°C utilizando tres diluyentes comerciales, indicados para la refrigeración de semen de otras especies monogástricas. Se realizaron diez colectas de semen de cada reproductora. Se mezclaron los eyaculados de cada conejo, obteniendo un pool de semen. Luego, el pool heterospérmico se dividió en tres tratamientos, según el diluyente utilizado: BotuDogⓇ (perros); BotuSemenⓇ (caballos); y Beltsville Thawing Solution - BTS® (cerdos). Las muestras fueron evaluadas para parámetros macroscópicos, microscópicos y funcionales en cuatro momentos: semen fresco (0 h) y refrigerado (16, 24 y 48 h). Los valores promedio de motilidad, vigor y defectos totales del semen fresco fueron 86,0%, 3,05 y 11,5%, respectivamente. Después del inicio del enfriamiento, se observó un comportamiento lineal decreciente para la motilidad y el vigor espermático del semen en función del tiempo, difiriendo (P < 0.05) entre tratamientos. Cabe destacar que el tratamiento diluido en BTS® mostró una reducción más pronunciada en el mantenimiento de los parámetros evaluados (P<0,05) a lo largo del tiempo, con los valores más bajos de motilidad espermática y vigor dentro de cada tiempo evaluado: 16h [20,6 ± 7,28 y 0,75 (0,375 -1.5)], 24h [12,0 ± 6,80 y 0 (0 - 1,0)] y 48h [3,0±2,13 y 0 (0 - 0,125)]. Después de 48 h de enfriamiento, BotuDogⓇ presentó el valor más alto de motilidad espermática (71,0 +2,08) y, junto con BotuSemenⓇ, presentó el valor más alto de vigor espermático (2,5). De acuerdo con los datos obtenidos, los diluyentes BotuDogⓇ y BotuSemenⓇ fueron eficientes en mantener parámetros espermáticos adecuados, mostrando buena viabilidad espermática hasta 48 horas después de la refrigeración a 5°C. A través de este trabajo se obtuvieron resultados inéditos respecto a parámetros espermáticos de semen de conejo refrigerado que pueden ser utilizados como valores de referencia para el manejo reproductivo y procesamiento de semen de estos animales, dada la falta de estas recomendaciones técnicas.

Cysticercosis in free-ranging agoutis (Dasyprocta leporina) in the Eastern Brazilian Amazon / Cisticercose em cutias (Dasyprocta leporina) silvestres na Amazônia Oriental brasileira

The study describes the occurrence of cysticercosis in liver of 22 wild agoutis (Dasyprocta leporina) in the Brazilian Amazon. The phylogenetic analysis and microscopic characteristics of metacestodes in liver tissue sections, associated with the geographic distribution of the intermediate hosts indicated that a possibly novel Taenia sp. metacestode caused the parasitism. Additionally, two cases of hepatic co-infection by Taenia sp., Calodium sp. and Echinococcus oligarthra were also observed among the analyzed animals. The results point to the need for a better understanding of hepatotropic parasites among wild rodents in the Brazilian Amazon.(AU)

O estudo descreve a ocorrência de cisticercose no fígado de 22 cutias (Dasyprocta leporina) silvestres da Amazônia brasileira. A análise filogenética e as características microscópicas dos metacestódeos em cortes histológicos de fígado, associadas à distribuição geográfica do hospedeiro intermediário, indicaram que, possivelmente, uma nova espécie de Taenia sp. Causou o parasitismo. Adicionalmente, dois casos de co-infecção por Taenia sp., Calodium sp. e Echinococcus oligarthra também foram observados entre os animais avaliados. Os resultados apontam para a necessidade de um melhor entendimento dos parasitas hepatotrópicos entre roedores selvagens da Amazônia brasileira.(AU)

Occurrence of IgG antibodies against Toxoplasma gondii, Neospora caninum, and Leptospira spp. in goats and sheep from an indigenous village in Pernambuco, Brazil / Ocorrência de anticorpos IgG contra Toxoplasma gondii, Neospora caninum e Leptospira spp. em caprinos e ovinos de uma aldeia indígena em Pernambuco, Brasil

This study aimed to determine the occurrence of anti-Toxoplasma gondii, Neospora caninum, and Leptospira spp. antibodies in sheep and goats raised in villages of the Xukuru do Ororubá indigenous community, Pernambuco, Brazil. A total of 180 serum samples from sheep and 108 serum samples from goats of both sexes and different ages were analyzed. For antibody research, indirect immunofluorescence antibody test (IFAT) were used for the protozoa T. gondii and N. caninum, and microscopic agglutination test (MAT) for Leptospira spp., with a cutoff titer of 1:64, 1:50 and 1:100, respectively. The frequency of anti-T. gondii antibodies was 16.6% (30/180) for sheep and 11.1% (12/108) for goats. The frequency of anti-N. caninum antibodies was 10.55% (19/180) for sheep, and 20.37% (22/108) for goats, while for Leptospira spp., 2.2% (4/180) of sheep and 1.85% (2/108) of goats reacted positively. The results obtained in this study are unprecedented in indigenous communities in the country and serve as an alert for monitoring goats and sheep from the Xukuru do Ororubá indigenous village regarding the occurrence and productive impact of infections by T. gondii, N. caninum, and Leptospira spp., in addition to the occurrence of the zoonosis toxoplasmosis and leptospirosis in the indigenous community.(AU)

Objetivou-se determinar a ocorrência de anticorpos anti-Toxoplasma gondii, Neospora caninum e Leptospira spp., em ovinos e caprinos criados em aldeias da comunidade indígena Xukuru do Ororubá, Pernambuco, Brasil. Foram analisadas 180 amostras de soro de ovinos e 108 amostras de soro de caprinos de ambos os sexos e diferentes idades. Para a pesquisa de anticorpos foi utilizada a técnica de Reação de Imunofluorescência indireta (RIFI), para os protozoários T. gondii e N. caninum e Aglutinação Microscópica (MAT) para Leptospira spp., com ponto de corte de 1:64, 1:50 e 1:100, respectivamente. A frequência de anticorpos anti-T gondii foi de 16,6% (30/180) em ovinos e 11,1% (12/108) em caprinos. A frequência de anticorpos anti-N. caninum foi de 10,55% (19/180) para ovinos e 20,37% (22/108) para caprinos, enquanto para Leptospira spp., 2,2% (4/180) dos ovinos e 1,85% (2/108) dos caprinos reagiram positivamente. Os resultados obtidos neste estudo são inéditos em comunidades indígenas do país e alertam para o monitoramento de caprinos e ovinos da aldeia indígena Xukuru do Ororubá, quanto à ocorrência e impacto produtivo de infecções por T. gondii, N. caninum e Leptospira spp., além da ocorrência de zoonoses como a toxoplasmose e leptospirose na comunidade indígena.(AU)

Cutaneous myxoma in broiler chicken detected during slaughtering

Background: A cutaneous or superficial myxoma is a benign neoplasm of dermal or subcutaneous fibroblast origin. Although rare, it has been previously described in several species, including poultry. It presents as a single node or soft mass with a gelatinous cut surface. Histopathological analysis is essential for diagnosis and to differentiate it from other mesenchymal neoplasms and inflammatory or degenerative processes. Microscopically, it consists of dermal or subcutaneous lobules of plump, stellate, or spindle-shaped, bland-looking cells embedded in a basophilic myxoid matrix. This report describes the pathological findings in a rare case of cutaneous myxoma in a 42-day-old broiler flock. Cases: During ante mortem inspection of a 42-day-old broiler flock at a slaughterhouse under the authority of the Federal Inspection Service (southern Brazil), nodular lesions or encrusted areas with yellow and black areas were observed in the head skin of less than 1% of animals. These lesions, approximately 0.5 cm in diameter, were observed on the comb, in the periocular skin region, and close to the animals' nostrils. During the breeding period, no health or epidemiological events were observed. Fragments of the lesions in the comb and periocular skin were collected and fixed in buffered 10% formalin. The samples were sent to the laboratory, routinely processed, and stained with hematoxylin and eosin and Alcian blue. Microscopically, the lesions consisted of irregular multifocal proliferation of connective tissue showing spindle cells with poorly demarcated borders and scarce cytoplasm in a slightly basophilic myxoid aspect matrix. The adjacent epidermis is compressed due to neoplastic proliferation. No areas of epithelial hyperplasia or inclusion bodies were observed. According to the pathologic description and considering its descriptive epidemiology, our main clinical suspicion was cutaneous fowl pox, a pathology characterized by the appearance of nodules in regions devoid of feathers. However, the microscopic changes observed were compatible with those described for cutaneous myxomas. In addition, the extracellular matrix was positive for Alcian Blue staining, which is an indicator of myxoma. In the present case, the SIF did not report the same macroscopic lesions in other flocks of the same origin. Discussion: Connective tissue tumors, including myxomas, occur considerably less frequently under field conditions. In addition, these neoplasms are more frequent in mature birds and are not usually described in broilers, as observed in this report. The cutaneous myxoma described in broilers is usually a sporadic neoplasm that does not cause zootechnical losses, as observed in the case report. Its etiology is unknown and has been associated with various factors, such as local trauma and foreign bodies. Some fragments of plant material from the breeding environment were microscopically detected in the encrusted areas, which may indicate previous trauma or a foreign body. Myxoma has been associated with avian leukosis virus (ALV) subgroup A, but SIF did not report the same macroscopic lesions in other flocks of the same breeder hen's origin in the present case. Furthermore, sporadic connective tissue tumors associated with the virus occur in mature chickens but not in broilers. Myxoma lesions should be considered in the differential diagnosis of other connective tissue tumors and infectious agents that cause lesions in the skin and subcutaneous tissue.

Malignant tumor of peripheral nerve sheath in the thoracolumbar region in a cat

Background: In veterinary medicine, peripheral nerve tumors (PNST) are classified from the cellular pattern and as benign and malignant (MPNST). The majorities of cases are benign and usually involve the skin and subcutaneous tissue of the head, neck, and limbs. Animals with MPTNS usually have spinal cord and spinal canal involvement and are also described in the small and large intestine, oral cavity, perirenal region, and urinary bladder. Treatment is performed according to the location of the neoplasm and metastasis is rarely described. The present article aims to report a case of a malignant tumor in the peripheral nerve sheath of a cat. Case: A 8-year-old domestic cat was presented with a history of progressive paresis in the pelvic limbs. On neurological evaluation, proprioceptive ataxia and proprioception deficit in the pelvic limbs were observed, suggestive of thoracolumbar injury. Blood work and serum biochemistry showed results within the reference for the species and the bi-directional immunochromatography test for FIV and FeLV were negative. The radiographs of the thoracolumbar segment did not show significant changes. Myelography with iodinated contrast and collection of cerebrospinal fluid (CSF) were performed. On myelography examination, iodinated contrast loss was found between the 6th and 7th thoracic vertebrae, while the CSF analysis was normal. The patient underwent laminectomy and durotomy for excision of a mass found dorsally to the spinal cord, with intradural location. The tumor was sent for histopathological examination. Microscopic analysis showed neoplastic proliferation of spindle-shaped cells with predominance of the Antoni A pattern. Immunohistochemical analysis demonstrated intense cytoplasmic staining of the neoplastic cells for vimentin and S100 protein. Based on these findings, the diagnosis of malignant peripheral nerve sheath tumor was made. Radiotherapy was suggested as adjuvant treatment after surgical procedure. However, the owner refuse to follow this recommendation due to cost restriction. The patient had a satisfactory clinical recovery, he has no longer presented proprioceptive ataxia or proprioception deficits in three months after the surgical procedure. Discussion: In the case described, the feline patient presented a history, clinical signs, and findings in the myelography exam related to MPNST in the region of the vertebral canal. This neoplasm usually involves the spinal canal or spinal cord, but has been described in other locations, such in the small and large intestine, oral cavity, perirenal region, and urinary bladder. Based on the findings of the physical examination and complementary exams, a surgical procedure was recommended. Then, laminectomy and dutoromy were performed to remove the mass. The treatment described in the literature in cases of MPTNS is surgery, which can be curative, but in some cases, the use of radiotherapy may be necessary. The histological patterns observed in this neoplasm are the Antoni A and B, in the case described the Antoni A pattern was observed. The definitive diagnosis was made by immunohistochemistry with vimentin and S100 protein. MPNST are few described in the thoracolumbar region in cats. The surgical treatment was curative, where improvement of the clinical signs could be observed three months after surgery. In addition, monitoring of patients is recommended to ascertain relapses and metastases.

Refrigeração de sêmen canino em meio diluente à base de água de coco (Cocos nucifera) natural / Canine semen refrigeration in a natural coconut water (Cocos nucifera)-based extender / Refrigeración de semen canino en un medio diluyente natural a base de agua de coco (Cocos nucifera)

A inseminação artificial em cadelas contribui para o melhoramento genético da espécie, previne algumas doenças sexualmente transmissíveis a partir da cópula e possibilita a reprodução de animais que não poderiam copular de forma natural, seja por motivos anatômicos, geográficos ou comportamentais. Todavia, nem sempre é possível utilizar o sêmen fresco, sendo assim necessário um diluente para resfriar e mantê-lo viável por determinado período. Diante disso, o objetivo com este trabalho foi analisar o sêmen canino diluído em água de coco e refrigerado, à 5 ºC em diferentes tempos. Foram utilizados cinco cães da raça Hounds do Brasil, realizando três colheitas de sêmen de cada animal, com intervalos de sete dias. Os ejaculados foram mantidos a temperatura de 37ºC e realizado análises macroscópicas (volume, cor, aspecto e odor) e microscópicas (motilidade, vigor, concentração e morfologia espermática). Em seguida, os ejaculados foram diluídos em água de coco natural a uma concentração de 200 milhões de espermatozoides/mL, e mantidos à temperatura de 5 °C, por até 72 horas. Nos intervalos de seis, doze, vinte e quatro, trinta e seis, quarenta e oito, e setenta e duas horas, as amostras foram avaliadas quanto a motilidade e vigor espermático. Os ejaculados frescos apresentaram em média volume de 6,2 mL, cor branca, aspecto aquoso a leitoso, odor "sui generis", motilidade espermática de 89,5 %, vigor espermático 4,3, concentração média de 418 x106espermatozoides/mL e 7,3% de alterações patológicas. Após o início do resfriamento à 5 ºC, os valores de motilidade e vigor diminuíram com o passar do tempo, sendo os menores valores encontrados após 48 e 72 horas. O diluente a água de coco in natura mostrou-se eficiente para refrigeração de sêmen canino, à 5 ºC, conservando-o por um período de até 36h após a colheita, conforme preconizado pelo Colégio Brasileiro de Reprodução Animal.(AU)

improvement of the species, prevents some sexually transmitted diseases through copulation and allows the reproduction of animals that could not copulate naturally, either for anatomical, geographic or behavioral reasons. However, it is not always possible to use fresh semen, thus requiring a diluent to cool and keep it viable for a certain period. Therefore, the objective of this work was to analyze canine semen diluted in coconut water and refrigerated at 5 ºC at different times. Five Brazilian Hounds were used, performing three semen collections from each animal, with intervals of seven days. The ejaculates were kept at a temperature of 37 ºC and macroscopic (volume, color, appearance and odor) and microscopic (motility, vigor, concentration and sperm morphology) analyzes were performed. Then, the ejaculates were diluted in natural coconut water at a concentration of 200 million sperm/mL, and kept at a temperature of 5 °C for up to 72 hours. At intervals of six, twelve, twenty-four, thirty-six, forty-eight, and seventy-two hours, samples were evaluated for sperm motility and vigor. Fresh ejaculates had an average volume of 6.2 mL, white color, watery to milky appearance, "sui generis" odor, 89.5% sperm motility, 4.3 sperm vigor, average concentration of 418 x106 spermatozoa/mL and 7.3%pathological changes. After the beginning of cooling at 5 °C, the values of motility and vigor decreased over time, with the lowest values found after 48 and 72 hours. The in natura coconut water extender proved to be efficient for cooling canine semen at5 ºC, keeping it for a period of up to 36 hours after harvest, as recommended by the Brazilian College of Animal Reproduction.(AU)

Artificial insemination in bitches contributes to the genetic La inseminación artificial en perras contribuye a la mejora genética de la especie, previene algunas enfermedades de transmisión sexual a través de la cópula y permite la reproducción de animales que no podrían copular de forma natural, ya sea por razones anatómicas, geográficas o de comportamiento. Sin embargo, no siempre es posible utilizar semen fresco, por lo que se requiere un diluyente para enfriarlo y mantenerlo viable durante un cierto período. Por tanto, el objetivo de este trabajo fue analizar semen canino diluido en agua de coco y refrigerado a 5 ºC en diferentes tiempos. Se utilizaron cinco sabuesos brasileños, realizándose tres colectas de semen de cada animal, con intervalos de siete días. Los eyaculados se mantuvieron a una temperatura de 37 ºC y se realizaron análisis macroscópicos (volumen, color, apariencia y olor) y microscópicos (motilidad, vigor, concentración y morfología espermática). Luego, los eyaculados se diluyeron en agua de coco natural a una concentración de 200 millones de espermatozoides/mL y se mantuvieron a una temperatura de 5 °C hasta por 72 horas. A intervalos de seis, doce, veinticuatro, treinta y seis, cuarenta y ocho y setenta y dos horas, se evaluó la motilidad y el vigor de los espermatozoides en las muestras. Los eyaculados frescos tuvieron un volumen promedio de 6,2 mL, color blanco, apariencia acuosa a lechosa, olor "sui generis", motilidad espermática de 89,5%, vigor espermático de 4,3, concentración promedio de 418 x106 espermatozoides/mL y cambios patológicos de 7,3%. Después del inicio del enfriamiento a 5 °C, los valores de motilidad y vigor disminuyeron con el tiempo, encontrándose los valores más bajos a las 48 y 72 horas. El diluyente de agua de coco in natura demostró ser eficaz para enfriar el semen canino a5 ºC, manteniéndolo por un período de hasta 36 horas después de la cosecha, según lo recomendado por el Colegio Brasileño de Reproducción Animal.(AU)

Adenocarcinoma pancreático em um gato / Pancreatic adenocarcinoma in a cat

Background: Despite being rare in domestic animals, pancreatic adenocarcinoma is the most common malignant tumor of the feline pancreas. Due non-specificity of clinical signs in cats and the late diagnosis of the neoplasm, it is necessary to understand this disease better, to contribute for the knowledge of its early recognition and treatment. Thus, this study aims to report a case of metastatic pancreatic ductal adenocarcinoma in a cat, focusing on the main clinical aspects, diagnosis, and prognosis of this disease, in addition to the description of the presentation of peritoneal carcinomatosis. Case: A 14-year-old male neutered mixed breed cat, was referred to the Feline Medicine Service (MedFel) of the Hospital de Clínicas Veterinárias (HCV) - Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, with a history of hyporexia, constipation and increased abdominal volume for 3 days, besides mild difficulty in locomotion and progressive weight loss in the last 6 months. On the physical examination, the patient was alert, with a body condition score of 6/9; muscle condition score 1/4 and moderate dehydration of 7%. Popliteal lymph nodes were enlarged, and abdominal distension was evident. Around 200 mL of a slightly cloudy, straw-yellow liquid were drained from the abdominal cavity. After draining the fluid, a new abdominal palpation was performed, and there were fecal retention and a palpable mass in the right hypogastric region. The result of the cytological analysis of the fluid was consistent with a protein-rich transudate, suggesting neoplastic effusion of epithelial origin. Hematological and biochemical changes included leukocytosis due to neutrophilia, monocytosis, lymphopenia, thrombocytosis and azotemia. On abdominal ultrasound, the patient had free fluid in the abdominal cavity, and the gallbladder had discreet of biliary sludge. The intestines showed some corrugated segments with other segments lacking definition of its layers, and without peristaltic movements, suggesting intestinal neoplasia. Pancreas and adrenals were not visualized. On the chest X-ray, moderate opacification of lung fields with a diffuse interstitial pattern was observed, suggesting lung metastasis. The patient presented an acute worsening of the clinical condition and the owner requested euthanasia. The patient was referred for necropsy and based on the macroscopic and microscopic changes, the post-mortem diagnosis was metastatic pancreatic ductal adenocarcinoma with peritoneal carcinomatosis. Discussion: The clinical presentation of cats with exocrine pancreatic neoplasia is nonspecific, as clinical signs are common to several diseases, such as anorexia, vomiting, abdominal pain, weight loss with normal appetite, jaundice, depression, and lethargy. Complementary blood tests also do not provide data that could lead to the suspicion of pancreatic neoplastic disease. In the present case, the diagnosis of metastatic pancreatic ductal adenocarcinoma with peritoneal carcinomatosis was only possible post mortem. The pancreas is a difficult organ to assess adequately using most diagnostic imaging methods, so histopathology is still the method of choice for differentiating pancreatic tissue comorbidities. Therefore, exploratory laparotomy should be instituted to provide tissue samples from the pancreas and its metastases for histopathological diagnosis, whenever ultrasound or other imaging methods indicate suspicious abdominal changes. The literature reports that less than 10% of affected cats treated with complete surgical removal of the mass and chemotherapy alone will survive more than a year, and the average time for untreated cats is only 6 days. The prognosis of this disease is bad and most cats are euthanized, due to rapid clinical worsening. Therefore, diagnosis is essential to determine an adequate prognosis in advanced cases and to support therapeutic decisions or euthanasia.

Study of the upper pole after subtotal splenectomy in rats

Purpose: To evaluate macro/microscopic viability of the upper pole (UP) in rats after 80 days of subtotal splenectomy preserving the upper pole (SSPUP). Methods: Twenty-five male Wistar rats were submitted to SSPUP. After 80 days, the rats were euthanized, and the remaining UP was evaluated macroscopically regarding appearance, color, consistency, length, width, thickness, and presence of fibrosis/necrosis; and microscopically regarding presence of red and white pulp, fibrosis/necrosis. Results: Two rats died during surgery and were removed from the statistical analysis. There was statistically significant increase in length and width between the pre and postoperative in the experimental group, with no significant difference in thickness. In the manipulation group, the macroscopic appearance of the spleen was normal in pre and postoperative, with viability preserved. In the experimental group, two UP of the spleen were not found during the second surgery. Macroscopically, it was observed absence of fibrosis and necrosis in all cases. Microscopically, the white and red pulp were intact in both groups. Two spleens of rats in the manipulation group presented areas with fibrosis and necrosis focus, which were not enough to be considered inviable. Conclusions: The UP of the spleen remained viable in 91.3% of the cases.

Ocorrência de perigos físicos em alimentos / Occurrence of physical hazards in food

A ocorrência de matérias estranhas com potencial perigo físico em alimentos é um grande desafio para a indústria alimentícia, e a análise microscópica é provavelmente o instrumento mais útil para detectá-las e identificá-las. Considerando a escassez de dados sobre os perigos físicos em produtos alimentícios no Brasil, o objetivo deste estudo foi descrever essa ocorrência nos produtos analisados no Núcleo de Morfologia e Microscopia do Centro de Alimentos do Laboratório Central do Instituto Adolfo Lutz (IAL), em São Paulo, no período de 2008 a 2020. Os resultados revelaram que, das 7.221 amostras de alimentos analisadas, 89 (1,2%) estavam em desacordo com a legislação em vigor, por conterem perigos físicos. De acordo com a categoria de alimentos, a ocorrência foi maior para bebidas (43%), seguida de cereais, farinha e farelo (22%). Quanto ao tipo de matéria estranha, os plásticos (duros e flexíveis) foram os mais frequentes (48%), seguidos dos metais (15%) entre as partículas perigosas detectadas nas amostras. A detecção e identificação de perigos físicos, por meio de análises microscópicas, contribuem para a segurança e qualidade dos produtos alimentícios oferecidos à população (AU).

The occurrence of physical hazards in food is a great challenge for the food industry, and microscopic analysis is probably the most useful instrument to detect and identify them. Considering the scarcity of data on physical hazards in food products in Brazil, the aim of this study was to describe this occurrence in the products analyzed at the Nucleus of Morphology and Microscopy of the Food Center of the Adolf Lutz Institute's Central Laboratory (IAL) in São Paulo, from 2008 to 2020. Results revealed that of the 7,221 analyzed food samples, 89 (1.2%) did not comply with the legislation due to the presence of physical hazards. According to the food category, the occurrence was higher in beverages (43%), followed by cereals, yeasts, flour and bran (22%). Regarding the type of foreign matter, among the dangerous particles detected, the most frequently found were plastics (48%) (hard and flexible), followed by metals (15%). The detection and identification of physical hazards by microscopic analysis contributes to the safety and quality of food products offered to the public. (AU)

Ovarian development of Xiphopenaeus kroyeri (Crustacea: Penaeidae) from Espírito Santo, southeastern Brazil / Desenvolvimento ovariano de Xiphopenaeus kroyeri (Crustacea: Penaeidae) no Espírito Santo, sudeste do Brasil

This study aimed to describe and characterize the stages of gonadal development of females of Xiphopenaeus kroyeri caught by artisanal fishers in Espírito Santo state, southeastern region of Brazil. All females (n= 1,831) were subjected to macroscopic and microscopic morphological analysis (n= 333) of the ovaries. From the morphology, coloration and degree of turgidity of the fresh ovary, macroscopic analysis determined five stages of gonadal development. The macroscopic analysis showed difficulties in differentiating the immature and spawning stages due to the similarity between the colors of the ovaries, which confirms the need to perform the macroscopic and histological analysis simultaneously for fisheries management studies. Microscopic observations allowed us to analyze the following six stages of cell development: oogonia, previtellogenic oocytes, primary vitellogenic oocytes, secondary vitellogenic oocytes, mature oocytes and atretic oocytes. From this, five stages of gonadal development were defined, i.e., immature, early development, advanced development, mature and spawned. The presence of peripheral bodies was not observed in this species. These results help to clarify and better understand the reproductive and population aspects of the Atlantic Seabob, which are fundamental for the establishment of management and conservation measures of this resource.

Este estudo teve como objetivo descrever e caracterizar os estágios de desenvolvimento gonadal de fêmeas de Xiphopenaeus kroyeri capturado por pescadores artesanais no Estado do Espírito Santo, região sudeste (Região sudeste = Rio de Janeiro, São Paulo, Minas Gerais e Espírito Santo) do Brasil. Todas as fêmeas (n= 1.831) foram submetidas à análise morfológica macroscópica e microscópica (n= 333) dos ovários. A partir da morfologia, coloração e grau de turgidez do ovário fresco, a análise macroscópica determinou cinco estágios de desenvolvimento gonadal. A análise macroscópica mostrou dificuldades em diferenciar os estágios imaturo e reprodutivo devido à semelhança entre as cores dos ovários, o que confirma a necessidade de realizar a análise macroscópica e histológica simultaneamente para estudos de manejo pesqueiro. As observações microscópicas permitiram analisar os seguintes seis estágios de desenvolvimento celular: oogônias, ovócitos prévitelogênicos, ovócitos em vitelogênese primária, ovócitos em vitelogênese secundária, ovócitos maduros e ovócitos atrésicos. A partir disso, foram definidos cinco estágios de desenvolvimento gonadal, ou seja, imaturo, desenvolvimento inicial, desenvolvimento avançado, maduro e desovado. A presença de corpos periféricos não foi observada nesta espécie. Estes resultados ajudam a esclarecer e compreender melhor os aspetos reprodutivos e populacionais do camarão do Atlântico, que são fundamentais para o estabelecimento de medidas de gestão e conservação deste recurso.

Evaluation of fecal smear methods for research on Cryptosporidium spp. oocysts in the feces of dairy calves / Avaliação dos métodos de esfregaço fecal para a pesquisa de oocistos de Cryptosporidium spp. nas fezes de bezerros da bovinocultura de leite

The objective of this study is to compare the direct fecal smear (DFS) and centrifugal sedimentation (CS) methods in the detection of Cryptosporidium spp. oocysts in fecal samples of dairy calves. One hundred and fourteen fecal samples were collected from calves aged up to six months from 10 dairy farms located in Palotina and Francisco Alves, Paraná, Brazil. The microscopic analysis revealed the presence of Cryptosporidium spp. oocysts in 51.75% (59/114) of the samples in both methods. In CS, 48.25% (55/114) of the samples were positive, while in DFS slides, only 6.14% (7/114) were positive. Only 4 samples were positive exclusively in DFS. To ensure that there were no false-negative results in the microscopic analysis, the 55 samples that were negative in both DFS and CS were selected for molecular analysis using the nested PCR (nPCR). Of these 55 samples, 24% (13/55) were positive and forwarded for sequencing part of the genome, which made it possible to identify C. parvum, C. bovis and C. ryanae. Besides the characterization of the Cryptosporidium species, it was possible to identify bacteria of the genus Acinetobacter interfering directly in the analyzed samples. The microscopic analysis also revealed higher sensitivity when CS was used to make the fecal smears. However, some samples that were negative in this technique had positive PCR results. Thus, molecular analysis is indicated to confirm cases of Cryptosporidium spp. Further studies are necessary to prove the specificities of the used primers since the results obtained in nPCR were positive for the protozoan but, when genetic sequencing was performed, Acinetobacter spp. was identified.(AU)

O objetivo deste trabalho foi comparar os métodos de esfregaço fecal direto (DFS) e centrífugo sedimentação (CS) para a pesquisa de oocistos de Cryptosporidium spp. em amostras fecais de bezerros leiteiros. Foram coletadas 114 amostras fecais de bezerros com até seis meses de idade, provenientes de dez propriedades leiteiras localizadas nos municípios de Palotina e Francisco Alves, Paraná. Por meio da análise microscópica, foi possível observar oocistos de Cryptosporidium spp. em 51,75% (59/114) das amostras. Pelo método da CS foi identificada positividade em 48,25% (55/114) das amostras, enquanto nas lâminas pelo método do esfregaço fecal direto (DFS), apenas 6,14% (7/114) foram positivas. Somente 4 amostras foram positivas exclusivamente no método do DFS. Para assegurar que na análise microscópica não houvesse resultados falso-negativos, as 55 amostras negativas para os dois métodos de confecção de lâminas (DFS e CS) foram selecionadas para análise molecular por meio da técnica de Nested PCR. Das 55 amostras submetidas a nPCR, 24% (13/55) apresentaram-se positivas e foram encaminhadas para o sequenciamento genético de uma porção do genoma, o qual possibilitou identificar as espécies de C. parvum, C. bovis e C. ryanae. Além da caracterização das espécies de Cryptosporidium, foi possível identificar a presença de bactérias do gênero Acinetobacter interferindo diretamente nas amostras analisadas. A análise microscópica revelou maior sensibilidade quando o método de CS foi usado para a confecção dos esfregaços fecais, entretanto, amostras negativas por meio dessa metodologia apresentaram resultados positivos na nPCR. Desta forma, para a confirmação dos casos de Cryptosporidium spp., é indicado a realização da análise molecular. Novos estudos são necessários para se comprovar a especificidade dos primers utilizados, uma vez que na nPCR o resultado obtido foi positivo, porém ao realizar o sequenciamento genético houve a identificação de Acinetobacter spp.(AU)

Diagnóstico de leptospira spp. en bovinos sacrificados en brasil / Diagnostic of leptospira spp. in slaughtered cattle from brazil / Diagnóstico de leptospira spp. em bovinos abatidos no brasil

La leptospirosis es considerada una zoonosis bacteriana de importancia en salud pública. Es común en áreas tropicales, especialmente en países en desarrollo con escasos recursos de salud y saneamiento. Este estudio evaluó la presencia de Leptospira spp. en bovinos sacrificados en un matadero de la región Centro-Oeste de São Paulo, Brasil, así como animales positivos identificados tanto en serología como por análisis molecular. Se investigaron muestras biológicas de sangre, hígado y riñones de 150 bovinos mediante la técnica de Prueba de Aglutinación Microscópica (MAT) y Reacción en Cadena de la Polimerasa convencional (cPCR). Los resultados serológicos mostraron que de los 150 animales, 71 (47,3%) fueron reactivos. Los resultados moleculares mostraron la presencia de Leptospira spp. en riñones de 21 (14%) animales, en hígado de cinco (3,3%) animales, en hígado y riñones en dos animales (1,3%) y en sangre, en un (0,7%) animal. Estos resultados indican una alerta sobre la salud de los bovinos de carne debido a la posibilidad de que estos animales sean fuente de infección y la importancia de la característica ocupacional de esta enfermedad. También se verificó la importancia de complementar las técnicas serológicas y moleculares.  

Leptospirosis is considered a bacterial zoonosis of public health importance. It is common in tropical areas, especially in developing countries with scarce health and sanitation resources. This study evaluated the presence of Leptospira spp. in slaughtered bovine in a slaughterhouse in the Midwest region of São Paulo, Brazil, as well as identified positive animals both in serology and by molecular analysis. Biological samples of blood, liver and kidneys from 150 cattle were investigated by the technique of Microscopic Agglutination Test (MAT) and conventional Polymerase Chain Reaction (cPCR). The serological results showed that of the 150 animals, 71 (47.3%) were reactive. The molecular results showed the presence of Leptospira spp. in kidneys of 21 (14%) animals, in liver of five (3.3%) animals, in liver and kidneys in two animals (1.3%) and in blood, in one (0.7%) animal. These results indicate a warning about the health of beef cattle due to the possibility of these animals being the source of infection and the importance of the occupational characteristic of this disease. It was also verified the importance of complementing serological and molecular techniques.

A leptospirose é considerada uma zoonose bacteriana de importância para a saúde pública. É comum em áreas tropicais, especialmente em países em desenvolvimento com escassos recursos de saúde e saneamento. Este estudo avaliou a presença de Leptospira spp. em bovinos abatidos em frigorífico da região Centro-Oeste de São Paulo, Brasil, e identificou animais positivos tanto por sorologia quanto em análise molecular. Amostras biológicas de sangue, fígado e rins de 150 bovinos foram investigadas pela técnica de Teste de Aglutinação Microscópica (MAT) e Reação em Cadeia da Polimerase convencional (cPCR). Os resultados sorológicos mostraram que dos 150 animais, 71 (47,3%) foram reativos. Os resultados moleculares mostraram a presença de Leptospira spp. nos rins de 21 animais (14%), no fígado de 5 animais (3,3%), no fígado e rins em 2 animais (1,3%) e no sangue em 1 animal (0,7%). Esses resultados indicam um alerta sobre a saúde dos bovinos de corte devido à possibilidade desses animais serem fonte de infecção e a importância da característica ocupacional desta doença. Verificou-se também a importância de complementar as técnicas sorológicas e moleculares.

Bothrops envenomation in dogs: local and systemic manifestations

Background: Snakebite envenoming is a condition that affects humans and domestic animals worldwide. Identification of the snake species involved in the envenomation is infrequent. Bothrops envenomation presents typical clinicopathological features. This report describes epidemiological, clinical, and pathological data of 2 cases of Bothrops envenomation in dogs, including the first case of Bothrops moojeni snake striking a domestic animal in Brazil. Cases: Case 1. A dog was witnessed to have a Bothrops moojeni snakebite on a farm. In the first 24 h, acute lameness, pain, diffuse swelling, focal bleeding at the left forelimb, and increased whole-blood clotting time were observed in the envenomed dog. Polyvalent antivenom was administered in addition to fluid therapy, analgesics, corticosteroids, and antibiotics. On the 5th day, the animal presented spontaneous bleeding at the wound site, thrombocytopenia, and increased whole-blood clotting time. An additional dose of polyvalent antivenom was administered, and local treatment at the snakebite site was initiated. After 13 days, the dog showed no clinical or laboratory changes and recovered entirely. Case 2. A mongrel dog was taken for a necropsy to determine the cause of death. Grossly, major findings included swelling in the nasal plane that extended to the neck and dissecting hemorrhage in the subcutaneous tissue and adjacent musculature. Hemorrhages were observed in the heart, parietal pleura, left forelimb, lumbar region, and perirenal tissue. Marked necrosis and disruption of small blood vessels and lymphatics within the deep dermis and subcutaneous tissue were the main microscopic findings close to the snakebite site. Additionally, degeneration and necrosis of muscle fibers and dissecting hemorrhage were observed in the head and neck tissues surrounding the snakebite site. Kidneys showed marked interstitial hemorrhage and acute tubular nephrosis. Discussion: Bothrops envenoming is characterized by local (hemorrhage, dermonecrosis, and myonecrosis) and systemic (coagulative disorders, systemic hemorrhage, and acute kidney injury) changes due to the effect of the main venom components such as phospholipase A2 and metalloproteinases. These changes are hallmarks for the bothropic envenomation, supporting the diagnosis in cases 1 and 2. In case 1, the dog developed a Bothrops moojeni snakebite envenomation, but the immediate treatment with antivenom allowed a favorable outcome. In case 2, gross and microscopic findings supported the presumptive diagnosis of fatal bothropic envenomation. A marked local reaction such as swelling, pain, bleeding, bruising, and tissue necrosis was observed in case 1. In case 2, the most significant local changes were swelling and edema at the head and neck, hemorrhage in the subcutaneous tissue, and adjacent musculature. Systemic effects were observed clinically as spontaneous bleeding, thrombocytopenia, increased whole-blood clotting time (Case 1), systemic hemorrhages, and acute tubular nephrosis (Case 2). A proper treatment probably prevented the development of acute renal failure in Case 1. Herein, we show the first case of accidental snakebite envenomation by B. moojeni in a dog in Brazil. Information is scarce on the identification of venomous snake species striking domestic animals. Fast detection of well-determined clinical and pathological findings of Bothrops envenomation is essential for a correct diagnosis, therapeutics, and a good prognosis, even in cases with an unknown history.