Outbreak of bovine tick-borne disease on a riverside property in the Western Amazonia: Case report / Surto de tristeza parasitária bovina em propriedade ribeirinha na Amazônia Ocidental: Relato de caso

The objective was to report an outbreak of tick-borne disease (TBD) on riverside property in the Western Amazon. The death of 25 Nellore cattle was reported on a rural property on the banks of the Purus River, state of Acre. The producer observed animals with staggering walking, drop in productivity, weight loss and evolution to death in approximately 30 days. Fifteen animals from the same batch were selected for clinical evaluation and the ear tip was punctured for hemoparasite research, in addition to blood collection for hematological, biochemical and molecular evaluation. The main laboratory findings were leukocytosis, thrombocytopenia, hypoproteinemia, elevated creatine kinase and reduced urea, creatinine and albumin, as well as visualization of forms suggestive of Anaplasma spp. in 13.33% of the samples. Through PCR, 20% positivity was observed for Anaplasmamarginale and 53.33% for Babesia sp. Hematological and biochemical changes, although highly suggestive, may suffer changes from other factors not related to TBD. Therefore, the presumptive identification of the etiological agent in the blood or confirmatory by molecular methods is essential in the diagnosis. Depending on the stage of the disease, low parasitemia occurs, making it difficult to see hemoparasites in blood smears. The Babesia sp. was the main agent of the outbreak of TBD in the population evaluated, which, when associated with early clinical and laboratory diagnosis, results in adequate therapeutic direction and prophylactic measures, promoting a balance between host, agent and vector.

Objetivou-se relatar um surto de tristeza parasitária bovina (TPB) em propriedade ribeirinha na Amazônia Ocidental. Foi notificado o óbito de 25 bovinos da raça Nelore, em uma propriedade rural às margens do rio Purus, estado do Acre. O produtor observou animais com andar cambaleante, queda na produtividade, perda de peso e evolução ao óbito em aproximadamente 30 dias. Quinze animais do mesmo lote foram selecionados para avaliação clínica e foi procedida a punção a ponta de orelha para pesquisa de hemoparasitos, além da coleta de sangue para avaliação hematológica, bioquímica e molecular. Os principais achados laboratoriais foram anemia, leucocitose, trombocitopenia, hipoproteinemia, elevação da creatina quinase e redução de ureia, creatinina e albumina, além da visualização de formas sugestivas de Anaplasma spp. em 13,33% das amostras. Por meio da PCR, foi observado 20% de positividade para Anaplasma marginale e 53,33% para Babesia sp. As alterações hematológicas e bioquímicas, embora bastante sugestivas, podem sofrer alterações de outros fatores não relacionados à TPB. Por isso, a identificação presuntiva do agente etiológico no sangue ou confirmatória por métodos moleculares é essencial no diagnóstico. A depender da fase da doença, ocorre baixa parasitemia, dificultando a visualização de hemoparasitos em esfregaços sanguíneos. A Babesia sp. foi o principal agente do surto de TPB na população avaliada, que, quando associado ao diagnóstico clínico e laboratorial precoce, resulta no direcionamento terapêutico adequado e medidas profiláticas, promovendo uma relação de equilíbrio entre hospedeiro, agente e vetor.

Prevalence and risk factors of Taenia hydatigena in dogs, pigs, and cattle in the Central Highlands of Vietnam.

Taenia hydatigena is a globally distributed canine tapeworm. The canine tapeworm results in economic impacts for farmers owing to organ condemnation. T. hydatigena utilizes dogs and other carnivores as definitive hosts while swine and ruminants serve as intermediate hosts. T. hydatigena is endemic in Vietnam; however, information on the prevalence and risk factors associated with infection is scarce. This cross-sectional study aims to identify the prevalence of T. hydatigena taeniasis in dogs and T. hydatigena cysticercosis in pigs and cattle. The risk factors associated with taeniasis in dogs were identified using a fixed effects logistic regression model and quantified using population attributable fractions. The prevalence of T. hydatigena taeniasis in dogs and T. hydatigena cysticercosis in pigs and cattle was 10.31% (95% CI 8.21 to 12.84%), 7.60% (95% CI 6.34 to 9.08%), and 11.11% (95% CI 7.63 to 11.81%), respectively; these levels were lower than those reported in other regions of Vietnam. The population attributable fraction of risk factors associated with T. hydatigena taeniasis in dogs for dogs living in proximity of an abattoir, those having access to raw pork or beef viscera, and those living in multi-dog households were 80%, 19%, and 7%, respectively. This current study identified and quantified the risk factors for taeniasis in dogs thus advocating for targeted community intervention programs to break the lifecycle of T. hydatigena in Dak Lak province.

Innate and humoral immune parameters at delivery in colostrum and calves from heifers experimentally infected with Neospora caninum.

Neospora caninum is a leading cause of abortion in cattle worldwide. The study of the immune response against N. caninum is critical to understand its epidemiology, pathogenesis, diagnosis and, ultimately, in preventing and controlling bovine neosporosis. Herein, we determined the gene expression of innate immune components endosomal RNA-sensing TLRs, BMAP28 cathelicidin, TNF-α and IL-10 and characterized the variation in both IgG ratio and avidity at delivery in N. caninum-infected heifers challenged at day 210 of gestation, colostrum and their calves. Increased BMAP28 expression was observed not only in colostrum but also in peripheral blood mononuclear cells (PBMC) and umbilical cord of calves from N. caninum-infected heifers in comparison with mock-infected control group. In addition, statistically significant decrease of TLR7 and IL-10 expression levels were observed in umbilical cord, suggesting an attempt to avoid an exacerbated immune response against the parasite. At delivery, serum and colostrum samples from infected group evidenced specific IgG anti-N. caninum. Infected heifers showed IgG1/IgG2 ratios <1 and high avidity specific IgG. As expected, colostrum samples of these animals exhibited a high IgG1 concentration and elevated avidity values. Three out of four calves from N. caninum-infected heifers had specific IgG with IgG1/IgG2 ratios>1 and lower avidity values before colostrum intake. Interestingly, both IgG1/IgG2 ratios and avidity values increased in seropositive calves after colostrum intake. Overall, this study provides novel information on neonatal immunity in congenitally infected calves, which is essential to understand how the immune pathways could be manipulated or immune components could be employed in order to improve protection against neosporosis.

Detection and genotyping of Giardia duodenalis from cattle and pigs in Hualien country, Eastern Taiwan.

BACKGROUND: Giardia duodenalis is a zoonotic protozoan parasite causing diarrhea through waterborne or fecal-oral infection. The cysts can live in the drinking water and cause pandemic diseases. In Taiwan, very little information is available regarding the epidemiology of G. duodenalis in domestic animals. METHODS: Fecal samples were collected from cattle (n = 156) and pigs (n = 141) in Hualien country, eastern Taiwan. Detection and genotyping were done by microscopy examination of fecal samples and amplification of the ß-giardin gene using nested PCR. RESULTS: The prevalence of G. duodenalis infection was 19.87% for cattle (31/156) and 4.26% for pigs (6/141). Using nested PCR, 30 infected samples found in cattle belonged to Assemblage E, and one sample belonged to Assemblage D. For pigs, four samples belonged to Assemblage E, one belonged to Assemblage D, and another one belonged to Assemblage A. In addition, these results showed that G. duodenalis Assemblage A was detected in pigs and may cause zoonotic transmission. CONCLUSION: This is the first epidemiological investigation of G. duodenalis infection in animals in Hualien, Taiwan. These results could provide epidemiological information for disease control and public health protection.

[Endemic status of schistosomiasis in People's Republic of China in 2019].

This report presented the endemic status of schistosomiasis in the People's Republic of China at a national level in 2019, and analyzed the data collected from the national schistosomiasis prevention and control system and 455 national schistosomiasis surveillance sites. Among the 12 provinces (municipality and autonomous region) endemic for schistosomiasis in China, Shanghai, Zhejiang, Fujian, Guangdong and Guangxi continued to consolidate the achievements of schistosomiasis elimination, Sichuan Province achieved transmission interruption, Jiangsu newly achieved the standard of transmission interruption and 5 provinces of Yunnan, Hubei, Anhui, Jiangxi and Hunan maintained transmission control by the end of 2019. There were 450 endemic counties (cities, districts) endemic for schistosomiasis, including 28 500 endemic villages covering 70 667 800 people at risk of infections. Among the 450 endemic counties (citis, districts), 66.89% (301/450), 28.44% (128/450) and 4.67% (21/450) kept the criteria of elimination, transmission interruption and transmission control of schistosomiasis, respectively. By the end of 2019, a total of 30 170 advanced schistosomiasis cases were documented in China. In 2019, a total of 12 090 712 individuals received inquiry examinations and 1 740 764 were positive; 5 158 369 individuals received serological tests and 89 753 were seropositive. A total of 327 475 individuals received stool examinations and 5 were positive, including one case of acute schistosomiasis. In 2019, snail survey was performed in 19 726 endemic villages in China and Oncomelania snails were found in 7 322 villages, accounting for 37.12% of all surveyed villages, with 6 villages with emerging snail habitats. Snail survey covered an area of 585 286.24 hm2 and 174 270.42 hm2 snail habitats were found, including emerging snail habitats of 64.20 hm2; however, no infected snails were identified. In 2019, a total of 605 965 bovines were raised in the schistosomiasis endemic areas of China, and 183 313 received serological examinations, with 1 176 positives detected, while 134 978 bovines received stool examinations, with 7 positives identified. In 2019, there were 28 557 patients with schistosomiasis receiving praziquantel chemotherapy, and expanded chemotherapy was given to 1 008 083 person-times; there were 7 bovines with schistosomiasis receiving praziquantel chemotherapy, and 296 053 herd-times expanded chemotherapy was given to bovines. In 2019, snail habitats at an area of 128 754.26 hm2 were given chemical treatment, and the actual area of chemical treatment was 69 605.55 hm2, while environmental improvements were performed in snail habitats covering an area of 2 847.00 hm2. Data from the 455 national schistosomiasis surveillance sites of China showed that the mean Schistosoma japonicum infection rates were both zero in humans and bovines in 2019, and no infected snails were found. The results demonstrate that the overall endemic situation of schistosomiasis remains at a lower infection level in China; however, there is still a risk of schistosomiasis transmission. To achieve the target set in the National Thirteenth Five-Year Plan for Schistosomiasis Control and consolidate the achievements of schistosomiasis control, precision control on schistosomiasis still needs to be reinforced in China.

Prevalence and molecular analysis of Sarcocystis infections in cattle in Northwest Iran and the first global report of S. gigantea in cattle.

Cattle are intermediate host for several species of Sarcocystis, including S. cruzi, S. hirsuta, and S. hominis with high prevalence worldwide. The present study aimed to determine the prevalence of Sarcocystis infection, species identification, and phylogenetic analysis of the parasite in cattle in Northwest Iran. The samples of diaphragm and esophagus from 290 cattle were collected from slaughterhouses in Northwest Iran and subjected to macroscopic, microscopic, and histopathology examinations, PCR-RFLP, sequencing and phylogenetic analyses. Tissue cysts of Sarcocystis spp. were detected in 92% of cattle by digestion and microscopic tests. Based on the PCR-RFLP and specific PCR, 87.9% and 1.03% of isolates were identified as S. cruzi, and S. hominis, respectively. Macrocyst was seen in a single sample that was identified as S. gigantea. The haplotype network exhibited the extension of the various haplotypes of S. cruzi between neighboring provinces in Northwest Iran. Heterogeneity analysis of S. cruzi 18S-rRNA sequences indicated genetic diversity among S. cruzi isolates (Haplotype diversity: 0.733-0.854) consisting 16 haplotypes; however, the nucleotide differences showed low diversity (0.01481 to 0.03351). Pair wise sequence distance matrix amongst S. cruzi sequences indicated an intra-species divergence of 0%-7.8% and identity of 92.6%-100%. Sarcocystis infection is highly prevalent in cattle in Northwest Iran, with the predominance of S. cruzi, and genetic variants of this species are unequivocally distributing in Northwest provinces. First global detection of S. gigantea in cattle reflects new insights of transmission dynamic and biology of this parasite in Iran.

Cysticercosis in Madagascar.

INTRODUCTION: Cysticercosis (CC) is the most common parasitic disease of the central nervous system. It is endemic in most developing countries where pigs are raised and consumed. An overview of all available data of this parasite in Madagascar is lacking. METHODOLOGY: We conducted a literature review, collecting information on published and available literature about cysticercosis in Madagascar between January 1st, 1990 and June 30th, 2020. RESULTS: Out of 858 publications; 61 were included, issued from peer-review indexed journals, non-indexed journals, books, Ministry reports and press releases. In Madagascar, porcine cysticercosis has been reported since 1901; human cysticercosis is highly prevalent with an overall estimated seroprevalence between 7 and 21%. Serological analysis is based on Enzyme-Linked Immunosorbent Assay (ELISA) and Enzyme-linked immunoelectrotransfer blot techniques (EITB) for confirmative testing. Neurocysticercosis (NCC) is the most common pattern of cysticercosis in Madagascar and it is reponsible for pediatric morbidity causing more than 50% of epilepsy cases. Though CT-Scan is now available and tends to be considered the gold standard for NCC diagnosis, it remains unaffordable for most Malagasy patients and implies the proposal of a diagnostic algorithm for physicians. CONCLUSIONS: Our review has revealed that human taeniasis and bovine cysticercosis is a considerable burden in Madagascar. A national control program has been developed aiming to decrease the seroprevalence rate from 16 to 10% in 2015. The aim of the country is now to implement a CC control and elimination program. Meanwhile, some massive cysticercosis screenings have been conducted in the capital Antananarivo to drive people's attention on this widespread infection.

Real-time PCR on skin biopsies for super-spreaders' detection in bovine besnoitiosis.

BACKGROUND: Bovine besnoitiosis, an emerging disease in Europe that can be transmitted by vectors, is caused by the apicomplexan Besnoitia besnoiti. Bovine besnoitiosis is difficult to control due to the complexity of its diagnosis in the acute stage of the disease, poor treatment success and chronically asymptomatic cattle acting as parasite reservoirs. When serological prevalence is low, detection and specific culling of seropositive cattle is feasible; however, economic considerations preclude this approach when serological prevalence is high. The aims of this study were to evaluate the accuracy of detection of super-spreaders in highly infected herds and to test their selective elimination as a new control strategy for bovine besnoitiosis. METHODS: Previous real-time PCR analyses performed on skin tissues from 160 asymptomatic animals sampled at slaughterhouses showed that the tail base was the best location to evaluate the dermal parasite DNA load. All seropositive animals (n = 518) from eight dairy or beef cattle farms facing a high serological prevalence of besnoitiosis were sampled at the tail base and their skin sample analysed by real-time PCR. A recommendation of rapid and selective culling of super-spreaders was formulated and provided to the cattle breeders. Subsequent serological monitoring of naïve animals was used to evaluate the interest of this control strategy over time. RESULTS: Among the 518 seropositive animals, a low proportion of individuals (14.5%) showed Cq values below 36, 17.8% had doubtful results (36 < Cq ≤ 40) and 67.8% had negative PCR results. These proportions were grossly similar on the eight farms, regardless of their production type (beef or dairy cattle), size, geographical location or history of besnoitiosis. Within two weeks of the biopsy, the rapid culling of super-spreaders was implemented on only three farms. The numbers of newly infected animals were lower on these farms compared to those where super-spreaders were maintained in the herd. CONCLUSIONS: Real-time PCR analyses performed on skin biopsies of seropositive cattle showed huge individual variabilities in parasite DNA load. The rapid culling of individuals considered as super-spreaders seems to be a new and encouraging strategy for bovine besnoitiosis control.

Diarreia em bezerros no sul do Rio Grande do Sul em um período de 34 anos / Diarrhea in calves in Southern Rio Grande do Sul in a period of 34 years

The objective of this work was to conduct a retrospective study of causes of diarrhea in calves in southern Rio Grande do Sul between 1980 and 2014. All necropsy protocols and materials received in the Regional Diagnostic Laboratory of the Veterinary School of the Federal University of Pelotas were reviewed. Ninety-four cases of calves with diarrhea were diagnosed, of these, 44.7% were caused by parasitic infections and 34.0% by bacterial infections. Parasites as cause of diarrhea was the most important diagnostic in southern Rio Grande do Sul and affected calves from three to 12 months of age. The occurrence of diarrhea in animals even infants may be associated with environmental factors and the form of management that these animals are kept. Bacterial diseases are also important causes of diarrhea in calves up to three months of age in the region.

O objetivo deste trabalho foi descrever os dados de um estudo retrospectivo nos registros de 34 anos atividade de um laboratório de diagnóstico sobre as causas de diarreia em bezerros na região sul do Rio Grande do Sul. Foram revisados os protocolos de necropsia e materiais desta espécie encaminhados ao Laboratório Regional de Diagnóstico da Faculdade de Veterinária da Universidade Federal de Pelotas. Foram diagnosticados 94 casos de bezerros com diarreia, destes, 44,7% foram de causas parasitárias e 34,0% de causas bacterianas. Diarreia de causa parasitária foi a mais importante na região sul do Rio Grande do Sul e afetou bezerros de três a 12 meses de idade. A ocorrência de diarreia em animais mesmo lactentes pode estar associada a fatores ambientais e a forma de manejo no qual esses animais são mantidos. As enfermidades bacterianas foram, também, importantes causas de diarreia em bezerros de até três meses de idade na região.

Diarreia em bezerros no sul do Rio Grande do Sul em um período de 34 anos / Diarrhea in calves in Southern Rio Grande do Sul in a period of 34 years

O objetivo deste trabalho foi descrever os dados de um estudo retrospectivo nos registros de 34 anos atividade de um laboratório de diagnóstico sobre as causas de diarreia em bezerros na região sul do Rio Grande do Sul. Foram revisados os protocolos de necropsia e materiais desta espécie encaminhados ao Laboratório Regional de Diagnóstico da Faculdade de Veterinária da Universidade Federal de Pelotas. Foram diagnosticados 94 casos de bezerros com diarreia, destes, 44,7% foram de causas parasitárias e 34,0% de causas bacterianas. Diarreia de causa parasitária foi a mais importante na região sul do Rio Grande do Sul e afetou bezerros de três a 12 meses de idade. A ocorrência de diarreia em animais mesmo lactentes pode estar associada a fatores ambientais e a forma de manejo no qual esses animais são mantidos. As enfermidades bacterianas foram, também, importantes causas de diarreia em bezerros de até três meses de idade na região.

The objective of this work was to describe the data from a retrospective study in the records of 34 years of activity in a diagnostic laboratory on the diarrhea causes of in calves in the southern region of Rio Grande do Sul. All necropsy protocols and materials received in the Regional Diagnostic Laboratory of the Veterinary School of the Federal University of Pelotas were reviewed. Ninety-four cases of calves with diarrhea were diagnosed, of these, 44.7% were caused by parasitic infections and 34.0% by bacterial infections. Parasites as cause of diarrhea was the most important diagnostic in southern Rio Grande do Sul and affected calves from three to 12 months of age. The occurrence of diarrhea in animals even infants may be associated with environmental factors and the form of management that these animals are kept. Bacterial diseases are also important causes of diarrhea in calves up to three months of age in the region.

Epidemiology of bovine cysticercosis and associated economic losses in the state of Rio Grande do Sul, Brazil.

Bovine cysticercosis (BCC) is the most frequently detected zoonosis in Brazilian slaughterhouses and is considered a problem for public health and beef production chain, requiring epidemiological studies focusing on evaluating its prevalence, spatial distribution, and economic losses in order to improve and adopt specific strategies for BCC control. Thus, this study focused to establish BCC prevalence and spatial distribution in the state of Rio Grande do Sul, Brazil, and estimate the economic losses for cattle farmers suppliers of one exporter slaughterhouse. A set of 70,591 bovine carcasses were postmortem inspected from 2019 to 2020, which came from 134 municipalities located in eight distinct regions in the state of Rio Grande do Sul. The prevalence found was 3.44% (C.I. 95% 3.30-3.57%), and animals infected with unviable cysticerci were more frequently detected (70.56%) than those carrying viable ones (29.44%). The most frequent destination of carcasses and viscera was non-export (65.48%), followed by freezing/salting (25.41%), heat treatment (8.74%), and rendering (0.37%), resulting in a total economic burden of at least US$ 167,868.53 for cattle farmers. Some regions had higher risk for BCC occurrence, such as Porto Alegre, Caxias do Sul, Santa Maria, Ijuí, and Passo Fundo (OR > 1, p < 0.05), respectively. These results highlight the need of adopting prophylactic measures, mainly in specific areas, in order to control BCC and reduce the economic losses for beef production chain.

ROS-mediated NLRP3 inflammasome activation participates in the response against Neospora caninum infection.

BACKGROUND: Neospora caninum is an obligate intracellular protozoan that causes neosporosis, N. caninum infection is a major cause of abortion in cattle worldwide. Currently, specific treatment for neosporosis is not available. The NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is a cytoplasmic protein complex that plays an important role in host defense against N. caninum infection, but the underlying mechanisms are poorly understood. METHODS: The reactive oxygen species (ROS) inhibitor and the ROS inducer, wild-type (WT) and NLRP3-deficient peritoneal macrophages or mice were used to investigate the role of ROS in NLRP3 inflammasome activation and controlling parasite burdens. ROS production, cell death and cell viability, production of inflammasome-mediated IL-1ß or IL-18, cleavage of caspase-1 and NLRP3 expression, as well as parasite burdens were detected. RESULTS: In vitro, N. caninum induced ROS generation in a dose-dependent manner in peritoneal macrophages. The pretreatment of ROS inhibitor N-acetyl-L-cysteine (NAC) significantly attenuated N. caninum-induced ROS production, LDH release, IL-1ß secretion and NLRP3 expression, whereas N. caninum proliferation was notably increased. In contrary, the ROS inducer pyrogallol (PG) significantly enhanced ROS production and NLRP3 inflammasome activity and decreased the parasite burden in N. caninum-infected peritoneal macrophages. NADPH-dependent ROS-mediated NLRP3 inflammasome activation induced by N. caninum can also be confirmed by using the NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI). However, the NAC or DPI pre-treatment or PG treatment did not significantly alter N. caninum-induced inflammasome activities and parasite proliferation in Nlrp3-/- peritoneal macrophages. In vivo, IL-18 releases in serum and parasite burdens in peritoneal exudate cells were significantly increased in PG-treated WT mice after infection with N. caninum; however, IL-18 productions and parasite burdens were not changed in PG-treated Nlrp3-/- mice. Furthermore, PG treatment in WT mice infected with N. caninum significantly decreased the mortality, weight loss and parasite burdens in tissues and histopathological lesions. CONCLUSIONS: Neospora caninum-induced NADPH-dependent ROS generation plays an important role in NLRP3 inflammasome activation and controlling parasites. The ROS inducer PG can control N. caninum infection mainly by promoting NLRP3 inflammasome activation. ROS-mediated NLRP3 inflammasome axis can be a potential therapeutic target for neosporosis.

Cysts and parasites in an abattoir in Northwest Ethiopia; an urgent call for action on "one health".

INTRODUCTION: Zoonotic parasitic infections such as echinococcosis affect cattle, sheep and goats by lowering quality of meat and hides as well as decreasing milk production. The burden of such diseases among humans is usually underestimated as they are difficult to diagnose. We used abattoir data to estimate the prevalence of zoonotic parasitic infections in animals. METHODOLOGY: Data from 2005-2018 was used from the registry of an abattoir in Northwest Ethiopia. Frequencies, proportions and trends over time were analyzed. Meat inspection was conducted by visualization, palpation and incision. RESULTS: A total of 58,787 animals were slaughtered in the abattoir during the study period. These included 51,956 (88 %) cattle, 5,890 (10%) sheep and 941 (2%) goats. The detected parasites included Echinococcus in 12,334/58,787 (21%) and Fasciola in 10,551/58,787 (18%) animals. Echinococcus infection was highest among goats (267/941, 28%), followed by cattle (11,591/51,956, 22%) and sheep (476/5,890, 8%). Fasciolosis was detected in 9,877/51,956 (19%) cattle and 178/941 (19%) goats. The number of animals slaughtered strongly decreased over time from 8,405 in 2006 to 1,605 in 2018. However, the proportion of parasitic infections remained high with some fluctuations over the study period. CONCLUSIONS: Echinococcosis and fasciolosis were very common with one out of five animals slaughtered infected. This is of public health concern and needs urgent multi-sectorial efforts from stakeholders at the national and regional level for control of these diseases. One health program approaches may warrant the control of transmission to humans.

Development of MALDI-TOF mass spectrometry for the identification of lice isolated from farm animals.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is now routinely used for the rapid identification of microorganisms isolated from clinical samples and has been recently successfully applied to the identification of arthropods. In the present study, this proteomics tool was used to identify lice collected from livestock and poultry in Algeria. The MALDI-TOF MS spectra of 408 adult specimens were measured for 14 species, including Bovicola bovis, B. ovis, B. caprae, Haematopinus eurysternus, Linognathus africanus, L. vituli, Solenopotes capillatus, Menacanthus stramineus, Menopon gallinae, Chelopistes meleagridis, Goniocotes gallinae, Goniodes gigas, Lipeurus caponis and laboratory reared Pediculus humanus corporis. Good quality spectra were obtained for 305 samples. Spectral analysis revealed intra-species reproducibility and inter-species specificity that were consistent with the morphological classification. A blind test of 248 specimens was performed against the in-lab database upgraded with new spectra and validated using molecular tools. With identification percentages ranging from 76% to 100% alongside high identification scores (mean = 2.115), this study proposes MALDI-TOF MS as an effective tool for discriminating lice species.

TITLE: Développement de la spectrométrie de masse MALDI-TOF MS pour l'identification de poux isolés d'animaux de ferme. ABSTRACT: La Spectrométrie de Masse à Temps de Vol par Désorption/Ionisation Laser Assistée après Matrice est maintenant utilisée pour l'identification rapide des microorganismes isolés à partir d'échantillons cliniques et a récemment été appliquée avec succès pour l'identification des arthropodes. Dans cette étude, cet outil protéomique a été utilisé pour identifier les poux prélevés sur le bétail et la volaille en Algérie. Les spectres MALDI-TOF MS de 408 spécimens adultes ont été mesurés pour 14 espèces, dont Bovicola bovis, B. ovis, B. caprae, Haematopinus eurysternus, Linognathus africanus, L. vituli, Solenopotes capillatus, Menacanthus stramineus, Menopon gallinae, Chelopistes meleagridis, Goniocotes gallinae, Goniodes gigas, Lipeurus caponis et Pediculus humanus corporis élevé en laboratoire. Des spectres de bonne qualité ont été obtenus pour 305 échantillons. L'analyse spectrale a révélé une reproductibilité intra-espèce et une spécificité inter-espèces qui concordaient avec la classification morphologique. Un test à l'aveugle de 248 échantillons a été effectué par rapport à la base de données de notre laboratoire mise à niveau avec de nouveaux spectres et validée à l'aide d'outils moléculaires. Avec des pourcentages d'identification allant de 76 à 100 % et des scores d'identification élevés (moyenne : 2,115), cette étude propose MALDI-TOF MS comme un outil efficace pour distinguer les espèces de poux.

Comprehensive Account on Prevalence and Characteristics of Hydatid Cysts in Livestock from Pakistan.

Pakistan is at intersection of hyperendemic regions for hydatidosis. Current study aimed to determine the prevalence of hydatid cysts and cyst characteristics in different intermediate hosts (sheep, goats, cattle and buffaloes) across the 4 provinces of Pakistan. A total of 991 sheep, 1,478 goats, 1,602 cattle and 1,343 buffaloes were examined for presence of hydatid cysts during 2 years (January 2016-December 2018). Differences in frequency of hydatidosis were observed with highest overall prevalence in buffaloes (11.9%) and sheep (11.5%). Highest prevalence and burden of infection were observed in older age animals (23.8%, 9.78±0.49) and females (26.5%, 12.53±0.67). Data for seasonal prevalence alluded to year-round presence of disease with non-significant statistical difference. Organ predilection indicated liver as the most preferred site of cyst localization followed mainly by lungs. An over-dispersion pattern was observed in all infected animals as majority of cysts belonged <10 cysts per infected host category. Highest percentage of fertile cysts was observed in liver of sheep. Interestingly, solitary form of cysts had higher fertility rate than multiple form. Amid lack of data and wide gap of knowledge, this study would try to fill up the lacunae regarding this neglected tropical disease. Extensive rearing of livestock, unregulated official slaughter and home slaughtering have played role in adaptability of E. granulosus in Pakistan.

Performance of the Dot-blot test method for detecting antibodies to Sarcocystis spp. in cattle / Desempenho do teste Dot-blot para detecção de anticorpos para Sarcocystis spp. em bovinos

Serological techniques can detect antibodies against Sarcocystis spp., Neospora caninum and Toxoplasma gondii antigens in single or mixed infections. Immunofluorescent antibody tests (IFAT) is considered the gold standard technique for Sarcocystosis diagnostic in cattle serum and a positive IFAT result reflects Sarcocystis spp. infection. Therefore, the aims of the present study were to compare IFAT and Dot-blot for sarcocystosis diagnostic in experimentally infected mice and to investigate serological cross-reactions with N. caninum and T. gondii in these methods. Mice (Mus musculus) were inoculated intraperitoneally with bradizoites of Sarcocystis spp. or tachyzoites of N. caninum or T. gondii. Serum samples were obtained and analyzed by IFAT and Dot-blot for the three protozoa. Serum from N. caninum and T. gondii experimentally infected mice were tested by IFAT and reacted only to N. caninum or T. gondii antigens, respectively. Specific antibodies against Sarcocystis spp. were present in all animals experimentally infected with this protozoan, with IFAT titers from 10 to 800. Serum samples from mice experimentally infected with Sarcocystis spp., N. caninum and T. gondii and tested by Dot-blot demonstrated no cross reaction between protozoa. A Dot-blot using Sarcocystis spp. antigen appears to be a good alternative to IFAT in the serological diagnosis of Sarcocystosis.(AU)

As técnicas sorológicas podem detectar anticorpos contra os antígenos de Sarcocystis spp., Neospora caninum e Toxoplasma gondii em infecções únicas ou mistas. O teste de anticorpos imunofluorescentes (IFAT) é considerado a técnica padrão-ouro para o diagnóstico de sarcocistose no soro de bovinos e um resultado positivo de IFAT reflete Sarcocystis spp. infecção. Portanto, os objetivos do presente estudo foram comparar IFAT e Dot-blot para diagnóstico de sarcocistose em camundongos infectados experimentalmente e investigar reações cruzadas sorológicas com N. caninum e T. gondii nesses métodos. Os camundongos (Mus musculus) foram inoculados intraperitonealmente com bradizoítos de Sarcocystis spp. ou taquizoítos de N. caninum ou T. gondii. As amostras de soro foram obtidas e analisadas por IFAT e Dot-blot para os três protozoários. O soro de N. caninum e T. gondii infectados experimentalmente foram testados por IFAT e reagiram apenas aos antígenos de N. caninum ou T. gondii, respectivamente. Anticorpos específicos contra Sarcocystis spp. estavam presentes em todos os animais experimentalmente infectados com este protozoário, com títulos de IFAT de 10 a 800. Amostras de soro de camundongos infectados experimentalmente com Sarcocystis spp., N. caninum e T. gondii e testadas por Dot-blot não demonstraram reação cruzada entre protozoários. Um Dot-blot usando Sarcocystis spp. O antígeno parece ser uma boa alternativa ao IFAT no diagnóstico sorológico da sarcocistose.(AU)

Pancreatic Hydatid Cyst Misdiagnosed as Mucinous Cystadenoma: CT and MRI Findings.

Isolated hydatid cysts of the pancreas are rare lesions, even in endemic regions. In this report, we present the case of a 76-year-old patient who was admitted to our clinic with a diagnosis of a cystic lesion in the tail of the pancreas. On preoperative computed tomography (CT) and magnetic resonance (MR) examination, the cyst was characterized as a mucinous cystadenoma. A laparoscopic distal pancreatectomy followed. A histopathological examination revealed a large hydatid cyst in the tail of the pancreas.

Morphological and molecular identification with frequency analysis of Calicophoron microbothrioides in central Chile / Identificação morfológica e molecular com análise de frequência de Calicophoron microbothrioides na região central do Chile

Abstract This research aimed to determine the presence of paramphistomids in cattle slaughtered in a slaughterhouse of the Ñuble Region of Chile, to identify flukes and to analyze the frequency of these parasites in the Maule, Ñuble, and Biobío administrative regions of Chile. Between October of 2016 and April of 2017, rumens of 494 cattle were examined for flukes in the forestomachs. Worms were identified morphologically and, in addition, molecular analysis of the internal transcriber spacer region 2 of the fluke's DNA was done and phylogenetic analyses were performed with Bayesian inference in 14 worms. The frequency was analyzed by locality (low- or highlands) and age. The overall frequency was 11.24%. The district with the highest frequency of presentation was Chillán Viejo (30.8%). Districts in the lowlands had similar frequencies to those in the mountain lands (p=0.1). The frequency of flukes was significantly higher in adult animals than in young ones (p<0.01). We obtained a 460 bp-length fragment of DNA that was identical to the sequences previously identified as Paramphistomum cervi and Calicophoron microbothrioides, and performed morphological analyses confirmed that our samples belonged to C. microbothrioides. This is the first published study of C. microbothrioides in Chile.

Resumo Este trabalho teve como objetivo determinar a presença de paramphistomídeos em bovinos abatidos em um matadouro da Região do Ñuble do Chile, para identificar parasitas e analisar a frequência desses parasitos nas regiões administrativas de Maule, Ñuble e Biobío, no Chile. Entre outubro de 2016 e abril de 2017, rúmens de 494 bovinos foram examinados à procura de vermes no pré-estômago. Os vermes foram identificados morfologicamente e, além disso, a análise molecular da região interna do espaçador do transcritor 2 do DNA e análises filogenéticas foram realizadas com inferência bayesiana em 14 vermes. A frequência foi analisada pela altitude da localidade (baixa ou alta) e idade. A frequência geral foi de 11,24%. O distrito com as maiores frequências de parasitismo foi Chillán Viejo (30,8%). Os distritos das terras baixas tinham frequências semelhantes às encontradas nas terras das montanhas (p=0,17). A frequência foi significativamente maior em animais adultos do que em jovens (p<0.01). Obtivemos um fragmento de DNA de 460 pb que era idêntico às sequências anteriores identificadas como Paramphistomum cervi e Calicophoron microbothrioides, e realizamos análises morfológicas que permitiram confirmar que nossas amostras pertenciam a C. microbothrioides. Este é o primeiro estudo publicado sobre C. microbothrioides no Chile.

Identification of antigen B (AgB) Gene polymorphism in cattle and sheep isolates of Echinococcus granulosus and investigation of effects on serological diagnosis.

Cystic Echinococcosis (CE) is a worldwide common helminth disease caused by the larval form of Echinococcus granulosus. The aim of this study is to determine the genetic differences between distinct isolates of E. granulosus obtained from cattle and sheep and determine the polymorphism of the AgB1 gene by DNA sequence analysis, as well as investigating its relationship with serological response using ELISA and Western Blot tests. For this aim, germinal membranes of hydatid cysts of 30 cattle and 30 sheep from the provinces of Elazig and Erzincan in Turkey and serum samples of these animals were collected. Following isolation of the total genomic DNA, the 12S rRNA gene of all isolates was amplified by PCR for genetic characterization, and the mt-CO1 gene region was examined by DNA sequence analysis. The gDNAs were then amplified by PCR using AgB1-specific primers, and genetic variation was investigated by DNA sequence analysis. At the final stage, all serum samples were analyzed by ELISA and Western Blot tests using a partially purified hydatid cyst fluid antigen. As a result, 114 (95%) of the 120 isolates were determined to be E. granulosus sensu stricto by using 12S rRNA-PCR. Subsequently, the DNA sequence analysis of the remaining 6 samples of the mt-CO1 gene revealed that all samples were E. granulosus sensu stricto. According to the DNA sequence analysis that followed, nucleotide changes in the AgB1 gene were observed in 13 (10.8%) of 120 samples. With this study, 9 (69.2%) out of 13 hydatid cysts in which polymorphism was detected by DNA sequence analysis in their AgB1 gene were found to be positive with ELISA, and 6 (46.1%) were found positive by WB. While 80 (74.7%) of 107 non-polymorphic samples in the AgB1 gene were found to be positive with ELISA, and 75 (70.9%) were positive with WB. As a result, variation in different ratios was determined in the AgB1 gene of E. granulosus sensu stricto, and it was determined that this had a partial effect on serological response.

Genotypes of Echinococcus isolated from domestic livestock in Kazakhstan.

The diversity and importance of Echinococcus species in domesticated animals in Kazakhstan are poorly understood. In this study, 17 cysts of Echinococcus were collected from cattle and a further 17 cysts from sheep. DNA was extracted from the individual cysts and used for polymerase chain reaction amplification of mitochondrial subunit 1 of the cox1 and nadh1 gene. Amplicon sequencing results revealed the presence of Echinococcus granulosus sensu stricto G1 in 15 cattle and 15 sheep, and G3 genotype from two cattle. Echinococcus canadensis (G6/G7 strain) was found in two cysts originating from sheep.