Natural association between bovine leukemia virus and reproductive infectious diseases

Background: Enzootic bovine leukosis (EBL) is a widespread infectious disease caused by the bovine leukemia virus (BLV), which results in immune system dysfunction. The resulting immunosuppression may lead to an increased prevalence of other diseases. Dairy cows infected have altered immune function associated with decreased milk production and shortened lifespan and decreased immune response to immunization. BLV infection, however, is often asymptomatic, so any connection between subclinical infection and common reproductive diseases remains unknown. This study aimed to describe the relationship between naturally occurring subclinical BLV and infectious reproductive diseases seroconversion in the field. Materials, Methods & Results: The diseases investigated included Bovine viral diarrhoea (BVD), Bovine alfaherpesvirus 1 (BoHV-1), Bovine gammaherpesvirus 4 (BoHV-4), Chlamydiosis, Leptospirosis, Brucellosis and Neosporosis in dairy cattle. Six hundred fifty-five sera samples from the northern and south-central regions of Uruguay, from asymptomatic female Holstein and Holstein crosses without a history of vaccination against reproductive diseases were processed using reference diagnostic methods (Seronautalization, ELISA, MAT, Rose Bengal Plate test). The seroprevalence of BLV was 20.0%. Seroprevalence of reproductive diseases BVD, BoHV-1 and BoHV-4 were 99.3%, 41.2% and 27.3% of the populations, respectively, and the total seroprevalence of Leptospirosis, bovine Neospora caninum and Chlamydiosis were 19.8%, 29.8% and 33.0% respectively. The results revealed positive associations between naturally contracted BLV and the presence of antibodies against BoHV-1 (P = 0.002), as well as between naturally contracted BLV and presence of antibodies against Leptospira spp. (P = 0.028). Discussion: BLV infection can impact innate and adaptive immune system cells and alter the proper functioning of uninfected cells. BLV infection may also induce changes in the complex balance of cytokine expression, cell proliferation, and programmed cell death in T- and B-lymphocytes, which is critical for immune competence and effective response to infectious challenges. The progression of BLV infection has a substantial effect on host defense mechanisms. Indeed, lowmagnitude serologic responses to a commercial foot-and-mouth disease vaccine and a J5 Escherichia coli vaccine have been observed. These results are supported by recent trial studies showing a reduced immune response to vaccination against BoHV-1 and Leptospira spp. in asymptomatic animals infected with BLV. These are 2 of the most prevalent infectious reproductive diseases in cattle worldwide, and our results provide evidence that a link between BLV and susceptibility to these diseases may exist. Although there is evidence of the co-occurrence of these diseases, it remains unknown whether there is a direct or indirect effect of BLV on infertility, embryonic loss, or abortion. Another possibility is that natural infection with these reproductive pathogens (BoHV-1, Leptospira, or others) promotes BLV expression, negatively affecting the farms where these pathogens are endemic. Considering the high seroprevalence of BLV in dairy herds in North and South America where the infection is endemic, it was explored BLV's role as an immunosuppressant by quantifying its co-occurrence with diseases that affect reproductive performance in breeding herds. Future work should clarify the role of BLV and the co-occurring pathogens in causing infertility or abortions.

Histology and histochemistry of the accessory gland of the female reproductive tract of Rhodnius neglectus Lent, 1954 (Hemiptera: Reduviidae) / Histologia e histoquímica das glândulas acessórias do sistema reprodutor feminino de Rhodnius neglectus Lent, 1954 (Hemiptera: Reduviidae)

Rhodnius neglectus is a wild triatomine, vector of the protozoan Trypanosoma cruzi, which causes Chagas' disease, and feeds on the blood of small mammals, being essential for its growth and reproduction. Accessory glands of the female reproductive tract are important in insect reproduction, but their anatomy and histology in R. neglectus are poorly studied. The aim of this work was to describe the histology and histochemistry of the accessory gland of the female reproductive tract of R. neglectus. The reproductive tract of five females of R. neglectus was dissected and the accessory glands transferred to Zamboni's fixative solution, dehydrated in a crescent series of ethanol, embedded in historesin, sectioned at 2 µm thick, stained with toluidine blue for histological analysis or mercury bromophenol blue for detection of total proteins. The accessory gland R. neglectus is tubular, without branches, opening in the dorsal region of the vagina and differing along its length in proximal and distal regions. In the proximal region, the gland is lined by the cuticle with a layer of columnar cells associated with muscle fibers. In the distal region of the gland, the epithelium has spherical secretory cells with terminal apparatus and conducting canaliculi opening in the lumen through pores in the cuticle. Proteins were identified in the gland lumen, terminal apparatus, nucleus and cytoplasm of secretory cells. The histology of the R. neglectus gland is similar to that found in other species of this genus, but with variations in the shape and size of its distal region.

Rhodnius neglectus é um triatomíneo silvestre, vetor do protozoário Trypanosoma cruzi, causador da doença de Chagas. Este inseto se alimenta do sangue de pequenos mamíferos, que é essencial para o seu crescimento e reprodução. As glândulas acessórias do sistema reprodutor feminino são importantes na reprodução de insetos, mas sua anatomia e histologia em R. neglectus são pouco conhecidas. O objetivo deste trabalho foi descrever a histologia e histoquímica da glândula acessória do aparelho reprodutor feminino de R. neglectus. O sistema reprodutor de cinco fêmeas de R. neglectus foi dissecado e as glândulas acessórias transferidas para solução fixadora de Zamboni, desidratadas em série crescente de etanol, embebidas em historesina, seccionadas com 2 µm de espessura, coradas com azul de toluidina para análise histológica ou submetidas ao teste de mercúrio-bromofenol para detecção de proteínas totais. O sistema reprodutor de R. neglectus tem uma glândula acessória tubular, sem ramificações, abrindo-se na região dorsal da vagina, sendo diferenciada em regiões proximal e distal. Na região proximal, a glândula é revestida internamente pela cutícula com uma camada de células colunares associadas a fibras musculares. Na região distal ocorrem células secretoras esféricas com aparato terminal e canalículos condutores que se abrem no lúmen da glândula através de poros na cutícula. O teste histoquímico revelou a presença de proteínas no lúmen da glândula e no aparato terminal, núcleo e citoplasma das células secretoras. A histologia da glândula de R. neglectus é semelhante à das espécies desse gênero, mas com variações na forma e no tamanho de sua região distal.

Immune response dynamics of recent and chronic small ruminant lentivirus infection in the male reproductive system / Dinâmica da resposta imunológica da infecção recente e crônica por lentivírus de pequenos ruminantes no sistema reprodutor de machos

The objective of this study was to analyze the immune responses of bucks to small ruminant lentivirus (SRLV) with a focus on the reproductive system of males with recent and chronic infection. A total of 12 bucks were selected, six seronegative and six seropositive with chronic natural infection for more than 18 months (chronic infection group). After selecting the animals, the six seronegative males were intravenously inoculated with caprine arthritis-encephalitis virus (CAEV)-Co viral strain at a titer of 10-5,6 TCID50/mL. After viral inoculation, this group was called the recent infection group and was monitored weekly with the chronically infected group for 180 days with blood serum and seminal plasma Western Blot (WB) analysis. Of the animals with chronic SRLV infection, 18.94% (50/264) showed anti-SRLV antibodies in at least one of the samples, and 81.06% (214/264) were negative. Anti-SRLV antibodies were detected in 27.27% (36/132) of the blood serum samples from this group, while 10.60% (14/132) were reactive in the seminal plasma WB test. The animals inoculated with CAEV-Co became seropositive after the third week of viral inoculation. In this group, 31.06% (41/132) of seminal plasma samples had anti-SRLV antibodies, and of these, 70.73% (29/41) coincided with blood serum results. Of the remaining 29.27% (12/41), the seminal plasma sample of only three animals (RIA2, RIA3, and RIA5) had anti-SRLV antibodies. One of the animals with a recent infection presented anti-SRLV antibodies only in seminal plasma samples, possibly due to virus compartmentalization. Intermittent viral shedding was observed in both biological samples, regardless of the infection stage. The immune response in bucks with recent SRLV infection is more significant than in chronically infected animals. Regardless of the stage of infection, there is a fluctuation in antibody levels, therefore, this creates a risk of false-negative samples when performing the diagnosis.

O objetivo desse estudo foi analisar a resposta imunológica aos lentivírus de pequenos ruminantes (LVPR) com enfoque no sistema reprodutor de machos com infecção recente e crônica. Para isso, foram selecionados 12 reprodutores caprinos, sendo seis soronegativos e seis soropositivos com infecção natural crônica há mais de 18 meses (grupo com infecção crônica). Após seleção dos animais, os seis machos soronegativos foram inoculados com a cepa viral do vírus da artrite encefalite caprina (CAEV)-Co, título 10-5,6 TCID50/mL, por via intravenosa. A partir da inoculação viral este agrupamento passou a ser denominado de grupo com infecção recente e juntamente com o grupo com infecção crônica foram acompanhados, semanalmente por 180 dias, com análise dos testes de Western Blot (WB) no soro sanguíneo e plasma seminal. Nos animais com infecção crônica para LVPR, 18,94% (50/264) apresentaram anticorpos anti-LVPR em pelo menos uma das distintas amostras, e 81,06% (214/264) tiveram resultados negativos. Das amostras de soro sanguíneo do referido grupo, em 27,27% (36/132) detectou-se anticorpos anti-LVPR, enquanto que no plasma seminal 10,60% (14/132) foram reagentes no teste de WB. Nos animais inoculados com o CAEV-Co, ocorreu a soroconversão após a terceira semana da inoculação viral. Nesse grupo, 31,06% (41/132) das amostras de plasma seminal tiveram anticorpos anti-LVPR, sendo que dessas 41, 70,73% (29/41) coincidiram com resultado das amostras de soro sanguíneo. Nos 29,27% (12/41) restante, houve a detecção somente no plasma seminal e eram amostras provenientes de três animais (AIR2, AIR3 e AIR5). Em um dos animais com infecção recente, só foi identificado anticorpos anti-LVPR em amostras de plasma seminal, possivelmente em função da compartimentalização do vírus. Intermitência viral foi observada em ambas as amostras biológicas, independentemente do estágio de infecção. Conclui-se que a resposta imunológica em reprodutores com infecção recente LVPR é mais acentuada do que em animais cronicamente infectados. E, independentemente do estágio da infecção há uma flutuação nos níveis de anticorpos, sendo, portanto, um fator de risco, em virtude da existência de amostras falso-negativo ao realizar o diagnóstico.

Patógenos transmitidos por vetores na reprodução canina / Vector-borne pathogens in canine reproduction

O artigo apresenta uma revisão sobre os efeitos reprodutivos da infecção pelos principais patógenos transmitidos por vetores que acometem cães, Leishmania spp., Babesia spp., Ehrlichia spp., Anaplasma spp. e Trypanosoma spp. São discutidas a patologia genital, fertilidade masculina e feminina, transmissão venérea, transmissão vertical, transmissão horizontal mãe-filho e sua relevância na epidemiologia das doenças.(AU)

The article presents a review of the reproductive effects of infection by main vector-borne pathogens of dogs, Leishmania spp., Babesia spp., Ehrlichia spp., Anaplasma spp. and Trypanosoma spp. Genital pathology, male and female fertility, venereal transmission, vertical transmission, horizontal mother-offspring transmission and their relevance to the epidemiology of the diseases are discussed.(AU)

Emergências reprodutivas de cadelas e gatas em um hospital veterinário universitário / Reproductives emergencies in female dogs and female cats treated in a teaching veterinary hospital

Emergências obstétricas são comuns na rotina clínica de cães e gatos. Dentre as emergências obstétricas uma das mais importantes é a distocia. De modo geral, a distocia ocorre quando há falha no momento do parto ou na expulsão do feto, sendo que a causa mais comum de distocia nas duas espécies é a inércia uterina primária. Outra emergência obstétrica de ocorrência frequente em cadelas e gatas é a piometra. Esta doença é uma infecção que ocorre no útero, causada por alterações hormonais na fase do diestro dos animais. As duas enfermidades (distocia e piometra) podem ter o envolvimento de anticoncepcionais, tanto nas cadelas como nas gatas. Em relação ao tratamento, na maioria das vezes, as distocias em cadelas e gatas são abordadas por meio da realização de cesarianas. Em piometra, a terapêutica mais efetiva é a realização de ovariohisterectomia. Neste estudo, 43 cadelas e 20 gatas com emergências obstétricas foram monitoradas durante 90 dias. Entre as cadelas, 29 (67,44%; 29/43) apresentaram piometra e 13 (30,23%; 13/43) distocia. Além disso, uma cadela apresentou prolapso vaginal. Entre as gatas, 13 apresentaram distocia (65%; 13/20) e sete (35%; 7/20) piometra. Algumas dessas emergências obstétricas foram relacionadas ao uso de anticoncepcional. Assim, os clínicos veterinários devem desencorajar esta prática e incentivar a castração das fêmeas.

Obstetric emergencies are common in the clinical routine of dogs and cats. One of the most important obstetric emergencies is dystocia. In general, dystocia occurs when there is a failure at the parturition time or expulsion of the fetus, and the most common cause of dystocia in both species is primary uterine inertia. Another frequently occurring obstetric emergency in bitches and cats is piometra. This disease is an infection that occurs in the uterus caused by hormonal changes in the diestrus phase of animals. In both diseases (dystocia and pyometra), contraceptives can be involved in female dogs and cats. Regarding the treatment, most of the time, dystocias in bitches and cats are addressed through cesarean sections. In pyometra, the most effective therapy is to perform ovariohysterectomy. In the present study, 43 female dogs and 20 female cats with emergency obstetric were monitored during 90 days. Among bitches, 29 (67.44%; 29/43) had pyometra, and 13 (30.23%; 13/43) showed dystocia. In addition, one female dog presented vaginal prolapse. Among the cats, 13 had dystocia (65%; 13/20) and seven (35%; 7/20) pyometra. Some of these obstetric emergencies were related to contraceptive use. Thus, veterinary clinicians must discourage this practice and encourage the neutering of females.

Emergências reprodutivas de cadelas e gatas em um Hospital Veterinário Universitário / Reproductives emergencies in female dogs and female cats treated in a Teaching Veterinary Hospital

Emergências reprodutivas, como as piometras e distocias, são frequentes na rotina clínica de cadelas e gatas. A distocia ocorre quando há falha no momento do parto ou na expulsão do feto e sua causa mais comum, nas duas espécies, é a inércia uterina primária. A piometra é uma infecção que ocorre no útero, causada por alterações hormonais na fase do diestro dos animais. As duas enfermidades (distocia e piometra) podem ter o envolvimento de anticoncepcionais, tanto nas cadelas como nas gatas. Na maioria das vezes, as distocias, em cadelas e gatas, é abordada por meio da realização de cesarianas, enquanto para a piometra, a terapêutica mais efetiva é a realização de ovariohisterectomia. Desta forma, tendo em vista a deficiência na quantificação de emergências reprodutivas obstétricas em cadelas e gatas diagnosticadas em serviços veterinários do Brasil, este trabalho teve como objetivo relatar os achados obtidos através do monitoramento, durante 90 dias, de 43 cadelas e 20 gatas com emergências reprodutivas. Dentre as cadelas, 29 (67,44%; 29/43) apresentaram piometra, 13 (30,23%; 13/43) distocia e 1 prolápso vaginal. Dentre as gatas, 13 (65%; 13/20) apresentaram distocia e sete (35%; 7/20) piometra. Algumas das emergências reprodutivas foram relacionadas ao uso de anticoncepcional. Assim, os clínicos veterinários devem desencorajar esta prática e incentivar a castração das fêmeas.

Reproductive emergencies, such as pyometras and dystocias, are frequent in the clinical routine of female dogs and cats. Dystocia occurs when there is a failure during delivery or expulsion of the fetus, and its most common cause, in both species, is primary uterine inertia. Pyometra is an infection that occurs in the uterus caused by hormonal changes in the diestrous phase of animals. In both diseases (dystocia and pyometra), contraceptives can be involved in both female dogs and cats. Most of the time, dystocias, in female dogs and cats, is addressed through cesarean sections, while in pyometra, the most effective therapy is to perform ovariohysterectomy. Thus, given the deficiency in the quantification of obstetric reproductive emergencies in female dogs and cats diagnosed in veterinary services in Brazil, this study aimed to report the findings obtained by monitoring 43 female dogs and 20 female cats with reproductive emergencies for 90 days. Among bitches, 29 (67.44%; 29/43) presented pyometra, 13 (30.23%; 13/43) showed dystocia, and 1 had vaginal prolapse. Among the cats, 13 presented dystocia (65%; 13/20), and 7 (35%; 7/20) showed pyometra. Some reproductive emergencies were related to contraceptive use. Thus, veterinary clinicians must discourage this practice and encourage the castration of females

Piometra em fêmeas domésticas: uma revisão / Pyometra in domestic females: a review / Piometra en hembras domesticas: una revision

Piometra é uma doença que afeta o útero de fêmeas causando infecção e inflamação com acúmulo de grande quantidade de exsudato purulento ou mucopurulento. Ocorre em todas as espécies domésticas, sendo mais comum em cadelas e vacas. Sua importância em cada espécie está relacionada ao número de casos e sua severidade, dentro de suas particularidades. O objetivo com esta revisão foi reunir estudos referentes à piometra em fêmeas domésticas, incluindo a epidemiologia, sinais clínicos, diagnóstico e tratamento nas principais espécies acometidas.

Pyometra is a disease that affects the uterus of females causing infection and inflammation with an accumulation of large amounts of purulent or mucopurulent exudate. It occurs in all domestic species, being more common in bitches and cows. Its importance in each species is related to the number of cases and its severity, within its particularities. The objective of this review was to gather studies referring to pyometra in domestic females, including epidemiology, clinical signs, diagnosis, and treatment in the main species affected.

La piometra es una enfermedad que afecta el útero de las hembras causando infección e inflamación com acumulación de grandes cantidades de exudado purulento o mucopurulento. Ocurre en todas las espécies domésticas, siendo más frecuente en perras y vacas. Su importancia en cada especie está relacionada con el número de casos y su severidad, dentro de sus particularidades. El objetivo de esta revisión fue reunir estudios sobre piometra en hembras domésticas, incluyendo epidemiología, signos clínicos, diagnóstico y tratamiento en las principales especies afectadas.

Leptospira spp. of the urinary tract of female carrier goats in semi-arid conditions

Background: Leptospirosis is an important infectious disease in goat farming, with a worldwide distribution. It is usually transmitted by rodents and the genital route, may cause reproductive losses, negatively impacting goat farming. The diagnosis lies on serological, molecular and isolation techniques. Considering the importance of this disease for small ruminants, this work aimed to evaluate the serological, molecular findings and isolation of pathogenic leptospires in the urinary tract (kidney and bladder tissues) of goats. Materials, Methods & Results: Thirty-four adult goats were used for slaughter. Renal samples (n = 34), bladder (n = 34), were collected for isolation of the agent and molecular detection of Leptospira sp. and blood samples (n = 34) for serological testing. The polymerase chain reaction (PCR) was used as a molecular test and the microscopic serum agglutination test (MAT) was used as a serological test. Samples with DNA amplification were subjected to genetic sequencing. The presence of Leptospira DNA was found in the tissues of 8 (23.4%) goats, and of these, only 2 were positive in PCR and MAT. There was a slight agreement between the PCR and MAT techniques (k = 0.150; P = 0.436). In 6 (17.6%) samples of renal tissue and 2 (5.8%) bladder samples, Leptospira DNA was detected. The genes in a kidney tissue sample were sequenced and demonstrated 99% similarity to Leptospira interrogans. Anti-Leptospira sp. were detected in 6 (17.6%) of the animals tested. Discussion: Serology identified 3 predominant serogroups: Icterohaemorrhagiae, Tarassovi and Autumnalis, serogroups that are related to the presence of rodents that coexist in rural environments. Autumnalis has been reported in small ruminants, raising the hypothesis that goats are adapted, becoming chronic carriers and possible maintenance hosts. The frequency obtained (17.6%) may be the result of the mixed breed pattern and rustic characteristics inherent to the goat species. Given the characteristics of the semi-arid region, such as low rainfall and high solar incidence, it is essential to use an adapted methodology, with a lower cut-off point (1:50), as the serological titer is an established relationship between the animal species, the level of exposure throughout its evolution and the region studied. Molecular findings and bacterial isolation reveal the agent's ability to colonize the urinary tract of goats. These data show the importance that urine has in the epidemiological chain, being able to transmit the agent through direct contact with this product or through contamination of soil and water. There was no statistical agreement between the diagnostic techniques used in this study, in this case, an association between PCR and MAT is recommended to obtain data with high sensitivity and specificity. A bladder sample was sequenced and showed 99% similarity to Leptospira interrogans. In the semiarid region, the most common form of leptospirosis spread is through the sale of animals in business fairs for breeding, rearing or slaughter, as well as sharing the same property with several breeders. The introduction of chronic and asymptomatic carriers on the properties represents a serious risk for the spread of the disease. The results show the presence of Leptospira spp. in semi-arid goat herds, having as risk factors the presence of rodents and intercropping. The association of MAT and PCR is necessary for a better diagnosis of the disease.

Genital infection by Aerococcus viridans in a captive african elephant (Loxodonta africana) / Infecção genital por Aerococcus viridans em um elefante africano em cativeiro (Loxodonta africana)

Aerococcus viridans is an emerging pathogen for humans and livestock animals, mainly associated with genitourinary infections cases. Its occurrence in wild mammals has never been reported. The aim of this study was to determine the etiological agent associated with clinical a case of a genital infection in a female African elephant (Loxodonta africana). Phylogenetic analysis and antimicrobial susceptibility profile of the isolate were also addressed. The animal presented frequent cases of genital infection with intermittent white secretion. Purulent secretion was sampled and submitted to bacteriological exam. The isolate obtained was thus identified by phenotypic and molecular methods as A. viridans and was found to be similar to human pathogenic isolates in BLASTn and phylogenetic analysis. The isolate was sensitive to almost all antimicrobials evaluated, presenting resistance to ciprofloxacin and norfloxacin. This is the first report of occurrence of A. viridans infection in the genital tract of an African elephant.(AU)

Aerococcus viridans é um patógeno emergente para seres humanos e animais de produção, principalmente associado a casos de infecções geniturinárias. Sua ocorrência em mamíferos selvagens nunca foi relatada. O objetivo deste estudo foi determinar o agente etiológico associado a um caso clínico de infecção genital em uma fêmea de elefante africano (Loxodonta africana). Análises filogenéticas e perfil de susceptibilidade antimicrobiana do isolado também foram avaliados. O animal apresentou casos frequentes de infecção genital com eliminação de secreção branca intermitente. A secreção purulenta foi coletada e submetida a exame bacteriológico. O isolado obtido foi identificado por métodos fenotípicos e moleculares como A. viridans e apresentou alta similaridade a isolados humanos patogênicos nas análises de BLASTn e filogenética. O isolado foi sensível a quase todos os antimicrobianos avaliados, apresentando resistência à ciprofloxacina e norfloxacina. Este é o primeiro relato de ocorrência de infecção por A. viridans no trato genital de elefante africano.(AU)

Condição microbiológica após antissepsia da genitália externa em éguas / Microbiological condition after antisepsis of reproductive tract in mares

A endometrite é a principal responsável pela diminuição nas taxas de fertilidade e recuperação embrionária em éguas e em parte, pode ser associada a contaminações por falhas na antissepsia das biotécnicas reprodutivas aplicadas à espécie. O objetivo deste trabalho foi avaliar os níveis de contaminação decorrentes de diferentes protocolos de antissepsia da genitália externa de éguas doadoras de embrião. Éguas sadias tiveram a vulva higienizada conforme um dos três grupos experimentais: (G1) sabão de côco; (G2) detergente neutro e (G3) clorexidina degermante 2%. Foi realizada a coleta de material da região da vulva, vestíbulo e útero com auxílio de swab estéril. As amostras foram processadas com objetivo de se determinar os principais microgranismos presentes, além da avaliação de Unidades Formadoras de Colônia (UFC). O G1 mostrou-se mais efetivo quanto à redução nas contagens de UFC em todas amostras. Por outro lado, G2 apresentou eficiência de 62%, sendo o tratamento menos eficiente na redução da contaminação das diferentes regiões avaliadas. A escolha adequada do agente sanitizante utilizado na higienização de períneo previamente a intervenções ginecológicas são de suma importância afim de se evitar contaminações ascendentes que levem a endometrites.(AU)

Endometritis is mainly responsible for the decrease in fertility and embryonic recovery rates in mares and, in part, it can be associated with contaminations due to failures in the antisepsis of reproductive biotechniques applied to the specie. The objective of this study was to evaluate the levels of contamination resulting from different protocols for antisepsis of the external genitalia of embryo donor mares. Healthy mares had their vulva sanitized according to one of the three experimental groups: (G1) coconut soap; (G2) neutral detergent and (G3) 2% chlorhexidine degerming. It was made the collection of material from the vulva, vestibule and uterus region with the aid of a sterile swab. The samples were processed in order to determine the main microorganisms present, in addition to the evaluation of Colony Forming Units (CFU). G1 was more effective in reducing CFU counts in all samples. On the other hand, G2 showed an efficiency of 62%, being the treatment less efficient in reducing contamination in the different regions evaluated. The proper choice of the sanitizing agent used to clean the perineum prior to gynecological interventions is of paramount importance in order to avoid ascending contamination that lead to endometritis.(AU)

Condição microbiológica após antissepsia da genitália externa em éguas / Microbiological condition after antisepsis of reproductive tract in mares

A endometrite é a principal responsável pela diminuição nas taxas de fertilidade e recuperação embrionária em éguas e em parte, pode ser associada a contaminações por falhas na antissepsia das biotécnicas reprodutivas aplicadas à espécie. O objetivo deste trabalho foi avaliar os níveis de contaminação decorrentes de diferentes protocolos de antissepsia da genitália externa de éguas doadoras de embrião. Éguas sadias tiveram a vulva higienizada conforme um dos três grupos experimentais: (G1) sabão de côco; (G2) detergente neutro e (G3) clorexidina degermante 2%. Foi realizada a coleta de material da região da vulva, vestíbulo e útero com auxílio de swab estéril. As amostras foram processadas com objetivo de se determinar os principais microgranismos presentes, além da avaliação de Unidades Formadoras de Colônia (UFC). O G1 mostrou-se mais efetivo quanto à redução nas contagens de UFC em todas amostras. Por outro lado, G2 apresentou eficiência de 62%, sendo o tratamento menos eficiente na redução da contaminação das diferentes regiões avaliadas. A escolha adequada do agente sanitizante utilizado na higienização de períneo previamente a intervenções ginecológicas são de suma importância afim de se evitar contaminações ascendentes que levem a endometrites.

Endometritis is mainly responsible for the decrease in fertility and embryonic recovery rates in mares and, in part, it can be associated with contaminations due to failures in the antisepsis of reproductive biotechniques applied to the specie. The objective of this study was to evaluate the levels of contamination resulting from different protocols for antisepsis of the external genitalia of embryo donor mares. Healthy mares had their vulva sanitized according to one of the three experimental groups: (G1) coconut soap; (G2) neutral detergent and (G3) 2% chlorhexidine degerming. It was made the collection of material from the vulva, vestibule and uterus region with the aid of a sterile swab. The samples were processed in order to determine the main microorganisms present, in addition to the evaluation of Colony Forming Units (CFU). G1 was more effective in reducing CFU counts in all samples. On the other hand, G2 showed an efficiency of 62%, being the treatment less efficient in reducing contamination in the different regions evaluated. The proper choice of the sanitizing agent used to clean the perineum prior to gynecological interventions is of paramount importance in order to avoid ascending contamination that lead to endometritis.

Selection and characterization of peptides mimetic to Campylobacter fetus subsp. venerealis using phage display / Seleção e caracterização de peptídeos miméticos a Campylobacter fetus subsp. venerealis usando phage display

Bovine genital campylobacteriosis (BGC) is a venereal and subclinical disease that affects the fertility of cattle herds, and it is caused by Campylobacter fetus subsp. venerealis . This study selected peptides mimetic to the BGC-causing agent from a phage library. Phage display is a technique that applies bacteriophage libraries that reveal peptides fused to the viral capsid in biological selections against target proteins. Biopannings were performed for biological selection in the phage library using rabbit hyperimmune serum and C. fetus subsp. venerealis protein extract. Five selected heptapeptides were considered mimetic to Cfv-NCTC 10354 based on the results of bioinformatics analysis and assays with hyperimmune serum and cervicovaginal mucus obtained from heifers. ALASLPL and LSYLFPP were the most reactive peptides and considered promising as possible mimetic immunogens for C. fetus subsp. venerealis.

Campilobacteriose Genital Bovina (CGB) é uma doença venérea e subclínica que causa problemas reprodutivos em rebanhos, causada por Campylobacter fetus subsp. venerealis. Este trabalho teve como objetivo selecionar peptídeos miméticos ao agente da CGB de uma biblioteca de fagos. Phage display é uma técnica que aplica bibliotecas de bacteriófagos que expõem peptídeos fusionados ao capsídeo viral em seleções biológicas contra proteínas alvo. Biopannings foram realizados para seleção biológica na biblioteca de fagos por meio de soro hiperimune de coelho e extrato proteico de C. fetus subsp. venerealis. Cinco heptapeptídeos selecionados foram considerados miméticos para Cfv-NCTC 10354 a partir de análises de bioinformática e ensaios com soro hiperimune e muco cérvico-vaginal de novilhas. ALASLPL e LSYLFPP foram os peptídeos mais reativos e considerados promissores como possíveis imunógenos miméticos para C. fetus subsp. venerealis.

Selection and characterization of peptides mimetic to Campylobacter fetus subsp. venerealis using phage display / Seleção e caracterização de peptídeos miméticos a Campylobacter fetus subsp. venerealis usando phage display

Bovine genital campylobacteriosis (BGC) is a venereal and subclinical disease that affects the fertility of cattle herds, and it is caused by Campylobacter fetus subsp. venerealis . This study selected peptides mimetic to the BGC-causing agent from a phage library. Phage display is a technique that applies bacteriophage libraries that reveal peptides fused to the viral capsid in biological selections against target proteins. Biopannings were performed for biological selection in the phage library using rabbit hyperimmune serum and C. fetus subsp. venerealis protein extract. Five selected heptapeptides were considered mimetic to Cfv-NCTC 10354 based on the results of bioinformatics analysis and assays with hyperimmune serum and cervicovaginal mucus obtained from heifers. ALASLPL and LSYLFPP were the most reactive peptides and considered promising as possible mimetic immunogens for C. fetus subsp. venerealis.(AU)

Campilobacteriose Genital Bovina (CGB) é uma doença venérea e subclínica que causa problemas reprodutivos em rebanhos, causada por Campylobacter fetus subsp. venerealis. Este trabalho teve como objetivo selecionar peptídeos miméticos ao agente da CGB de uma biblioteca de fagos. Phage display é uma técnica que aplica bibliotecas de bacteriófagos que expõem peptídeos fusionados ao capsídeo viral em seleções biológicas contra proteínas alvo. Biopannings foram realizados para seleção biológica na biblioteca de fagos por meio de soro hiperimune de coelho e extrato proteico de C. fetus subsp. venerealis. Cinco heptapeptídeos selecionados foram considerados miméticos para Cfv-NCTC 10354 a partir de análises de bioinformática e ensaios com soro hiperimune e muco cérvico-vaginal de novilhas. ALASLPL e LSYLFPP foram os peptídeos mais reativos e considerados promissores como possíveis imunógenos miméticos para C. fetus subsp. venerealis.(AU)

Transmissible Venereal Tumor associated with cutaneous metastasis and leishmaniasis in a bitch

Background: The Transmissible Venereal Tumor (TVT) is a round cell neoplasia, of contagious nature, common in the canine species, which mainly affects external genitalia. Despite metastasis being uncommon, the extragenital involvement canoccur via the lymphatic or hematogenous route to regional lymph nodes, skin, subcutaneous tissue, nasal and oral mucosa,as well as the central nervous system. When the location of the tumor is genital the clinical diagnosis can be conclusive,however if it presents extragenital forms, cytological or histopathological studies are necessary for the confirmation. Thiswork describes a case of TVT with cutaneous metastasis in a female dog with leishmaniasis.Case: A 3-year-old crossbred female dog was attended at the Small Animal Medical Clinic (CMPA) of the UniversityVeterinary Hospital (HVU) of the Federal University of West Bahia (UFOB), Barra Multidisciplinary Center. This neuteredbitch dewormed, never vaccinated, rescued from the streets at the age of two and a half, had a prior history of ehrlichiosisand pyometra. The owner reported apathy, anorexia, cachexia, depression and sternal decubitus of the animal, as wellas episodes of vomiting and recurrent fever. During the physical examination were observed respiratory and heart rateswithin normality ranges, with predominance of slightly pale mucous membranes, reactive left prescapular lymph node andsubcutaneous nodular mass, not adhered to the musculature, located in the medial portion of the thirteenth left rib. Werealso evidenced onychogryphosis, opaque, shineless and brittle fur, with pruritic and lichenified exfoliative dermatitis inthe left scapular region, ear tips and snout, presence of a discreet quantity of brownish-colored vulvar discharge with aputrid odor. In the cytological examination, was verified the presence of cells with characteristics of TVT located in thegenital mucosa, developing atypical...

Transmissible Venereal Tumor associated with cutaneous metastasis and leishmaniasis in a bitch

Background: The Transmissible Venereal Tumor (TVT) is a round cell neoplasia, of contagious nature, common in the canine species, which mainly affects external genitalia. Despite metastasis being uncommon, the extragenital involvement canoccur via the lymphatic or hematogenous route to regional lymph nodes, skin, subcutaneous tissue, nasal and oral mucosa,as well as the central nervous system. When the location of the tumor is genital the clinical diagnosis can be conclusive,however if it presents extragenital forms, cytological or histopathological studies are necessary for the confirmation. Thiswork describes a case of TVT with cutaneous metastasis in a female dog with leishmaniasis.Case: A 3-year-old crossbred female dog was attended at the Small Animal Medical Clinic (CMPA) of the UniversityVeterinary Hospital (HVU) of the Federal University of West Bahia (UFOB), Barra Multidisciplinary Center. This neuteredbitch dewormed, never vaccinated, rescued from the streets at the age of two and a half, had a prior history of ehrlichiosisand pyometra. The owner reported apathy, anorexia, cachexia, depression and sternal decubitus of the animal, as wellas episodes of vomiting and recurrent fever. During the physical examination were observed respiratory and heart rateswithin normality ranges, with predominance of slightly pale mucous membranes, reactive left prescapular lymph node andsubcutaneous nodular mass, not adhered to the musculature, located in the medial portion of the thirteenth left rib. Werealso evidenced onychogryphosis, opaque, shineless and brittle fur, with pruritic and lichenified exfoliative dermatitis inthe left scapular region, ear tips and snout, presence of a discreet quantity of brownish-colored vulvar discharge with aputrid odor. In the cytological examination, was verified the presence of cells with characteristics of TVT located in thegenital mucosa, developing atypical...(AU)

Campylobacter fetus in abomasal fluid from spontaneously aborted bovine and ovine fetuses

Pregnancy losses are a major concern in livestock industry due to their economic impact on producers. Campylobacter fetus subspecies fetus (Cff) and C. fetus subspecies venerealis (Cfv) are directly related to reproductive failures in ruminants. Cff colonizes the gastrointestinal tract of a wide range of hosts leading to abortion, while Cfv is restricted to genital tract being generally associated to infertility in bovine. Considering the great economic losses related to campylobacteriosis in cattle and ovine herds, this study aims to investigate the occurrence of C. fetus, considering Cff and Cfv subspecies, in bovine and ovine spontaneously aborted fetuses in state of Rio Grande do Sul, Brazil. In this study, samples of abomasal fluid collected from 30 spontaneously aborted bovine (n = 18) and ovine (n = 12) fetuses were investigated for the detection of Campylobacter fetus throughout conventional PCR. Positive fetuses for C. fetus presence were further analyzed by molecular assays for Cff and Cfv detection, in order to determine subspecies identification. When available, samples of the main organs of the thoracic and abdominal cavities, as well as the brain, skeletal muscle, eyelid, skin, and placenta were collected for further histopathological analyses and bacterial culture, aiming to assess the presence of infection lesions and pathogens in those sites, respectively. Additionally, RT-qPCR assays were also performed for the detection of ruminant pestivirus, in order to detect bovine viral diarrhea cases. Throughout the present methodology, C. fetus was detected in the abomasal fluid samples of 2 bovine fetuses, being both identified as Cfv subspecies by PCR. Histopathological analyses demonstrated that macroscopic and microscopic changes found in the Cfv-positive animals were not either specific or directly related to Campylobacter infections. Moreover, no significant bacterial growth was observed in microbiological culture from the collected tissues, and both fetuses were negative for ruminant pestivirus. Differently, there was no detection of C. fetus in any of the analyzed ovine fetuses. Considering that abortion diagnosis rates reported in cattle and sheep industry are highly variable among the published studies, and that abortion diagnoses are commonly inconclusive due to difficulties in sampling methodology and inadequate identification of the pathogen involved, it is important to investigate the etiological causes of abortion the herds for better understanding the causes of pregnancy issues and monitoring their occurrence. In addition, the absence of pathognomonic lesions in the tissues investigated in the histopathological analyses observed in this study strongly suggests that well-known etiological agents commonly associated to abortion, such as Leptospira spp., Toxoplasma spp., Chlamydia spp. and Neospora caninum, are unlikely to be the cause of infection of the analyzed fetuses. Taking this into account, the presence of C. fetus in the abomasal fluid samples from two bovine fetuses demonstrated in the present study suggests the possible association of Cfv not only with infertility, but also with cases of bovine abortion, highlighting the importance of investigating unusual causal agents of abortions in sheep and cattle. Overall, an adequate diagnosis is essential for establishing better prevention strategies to avoid the circulation of abortion-related infectious agents in the herds.(AU)

Morphology and molecular phylogeny of Heterobothrium lamothei (Monogenea: Diclidophoridae), a gill parasite infecting the tiger puffer fish Lagocephalus sceleratus (Tetraodontidae) / Morfologia e filogenia molecular de Heterobothrium lamothei (Monogenea: Diclidophoridae), um parasita das guelras que infecta o peixe tigre pufferLagocephalus sceleratus (Tetraodontidae)

Members of the family Diclidophoridae are potentially dangerous species for the puffer fish aquaculture worldwide. They are parasitic polyopisthocotyleans, with a posterior haptor equipped with clamps for attachment to the host's surface, allowing the worm to resist the flow of water to maintain its position on gills. The anterior body of the worm is deformable, allows the worm to feed on blood sucked from fish gills. The present study is the first description of a Heterobothrium species from the gills of the tiger puffer Lagocephalus sceleratus (Tetraodontidae) from the coasts of the Arabian Gulf at Jubail, Saudi Arabia morphologically by light microscopy as well as by molecular analysis of the parasite partial 28S rRNA through multiple sequence alignments and phylogeny by maximum likelihood analysis which is provided for the first time for the described species. Seventeen tiger puffer fish were captured alive from marine water off Saudi Arabia; gills were separated and further examined for parasitic infection. Nine fish were found infected with a monogenean parasite which was robust, equipped by two buccal organs at the tapered anterior end; the posterior haptor was rectangular with four symmetrically arranged clamps, with no isthmus. Marginal hooks absent. Ovary elongated, U-shaped, testes numerous, irregularly shaped and extended from the posterior part of the ovary to the anterior margin of the haptor. Copulatory organ muscular, as a spherical cup armed with 12 to 15 genital hooks. The molecular analysis of the parasite 28s rRNA and phylogeny revealed a percentage of identities between 87.47-89.09%, with Diclidophoridae species within the monophyletic clade of Mazocraeidea where a maximum percentage of 89.09% were obtained for the morphologically different sister taxon H. okamotoi. The results obtained from molecular analysis are consistent with the conclusions drawn from morphological classification where that the parasite recorded was morphologically similar to H. lamothei which was not characterized by molecular analysis before. The recovered sequences were deposited into the GeneBank under accession number MT322610.(AU)

Os membros da família Diclidophoridae são espécies potencialmente perigosas para a aquicultura de peixes puffer em todo o mundo. Eles são parasitas poliopisthocotyleans, com uma hélice posterior equipada com pinças para fixação na superfície do hospedeiro, permitindo que o verme resista ao fluxo de água para manter sua posição nas brânquias. O corpo anterior do verme é deformável, e permite que o verme se alimente de sangue sugado das guelras dos peixes. O presente estudo é a primeira descrição de uma espécie Heterobothrium das guelras do tigre Lagocephalus sceleratus (Tetraodontidae) das costas do Golfo Arábico em Jubail, Arábia Saudita, usando morfologia por microscopia leve, bem como análise molecular do rRNA parcial do parasita 28S através de alinhamentos de sequências múltiplas e filogenia por análise de máxima verossimilhança que é fornecida pela primeira vez para as espécies descritas. Dezessete peixes tigre puffer foram capturados vivos da água marinha da Arábia Saudita; as brânquias foram separadas e mais tarde examinadas para detecção de infecção parasitária. Nove peixes foram encontrados infectados por um parasita monogênio robusto, equipado por dois órgãos bucais na extremidade anterior afilada; o hortetor posterior era retangular com quatro pinças dispostas simetricamente, sem istmo. Ausência de ganchos marginais. Ovário alongado, em forma de U, testículos numerosos, de forma irregular e estendido desde a parte posterior do ovário até a margem anterior do hortelino. Órgão copulatório muscular, como um copo esférico armado com 12 a 15 ganchos genitais. A análise molecular do parasita 28s rRNA e filogenia revelou uma porcentagem de identidades entre 87,47-89,09%, com espécies Diclidophoridae dentro do clade monofilético de Mazocraeidea onde uma porcentagem máxima de 89,09% foi obtida para o táxon-irmão morfologicamente diferente H. okamotoi. Os resultados obtidos da análise molecular são consistentes com as conclusões tiradas da classificação morfológica onde o parasita registrado era morfologicamente semelhante ao H. lamothei que não era caracterizado pela análise molecular antes. As sequências recuperadas foram depositadas no GeneBank sob o número de acesso MT322610.(AU)

New approaches to studying morphological details of intramolluscan stages of Angiostrongylus vasorum / Novas abordagens para estudo morfológico dos estágios intramoluscos de Angiostrongylus vasorum

Angiostrongylus vasorum is a pulmonary artery parasite of domestic and wild canid. On molluscs, intermediate host, first stage larvae (L1) are found after the first day of infection, in the 8th L2 and in the 30 th L3. It was evaluated L1, L2 and L3 recovered by Baermann technique from Achatina fulica infected with 1000 L1. Fifty larvae/stage were incubated with antibodies anti-β-tubulin, anti-α-tubulin, anti- α-actin, anti-β-actin and anti-collagen, and then with Alexa 633. Fifty larvae/stage were observed with picrosirius red and Oil Red O. It was also observed in the anterior region of L1 the beginning of the chitinous stems development, in the initial portion of the intestine and genital primordium. In L2 anterior region, the papillae, chitinous canes juxtaposed to the mouth and intestines bigger than L1. The L3 musculature is well defined, next to the chitinous stems, there are two round distally arranged from each other. It was observed the whole extension of the intestine genital primordium and intense cellularity in the L3 distal portion. With the picrosirius red the L1, L2 and L3 musculature could be observed, as the nerve ganglia on L3. Oil Red O revealed that L1, L2 and L3 store energy on lipid droplets.(AU)

Angiostrongylus vasorum é um parasito de artérias pulmonares dos canídeos domésticos e silvestres. Nos moluscos, hospedeiros intermediários, encontram-se no primeiro dia após a infecção, larvas de primeiro estágio (L1), ao 8° L2 e ao 30° L3. Avaliou-se L1, L2 e L3 recuperadas pela técnica de Baermann de Achatina fulica infectada com 1.000 L1. Incubou-se 50 larvas/estádio com anticorpos anti-β-tubulina, anti-α-tubulina, anti-β-actina e anti-colágeno e, em seguida, com anticorpo Alexa 633. Observaram-se também 50 larvas/estádio com picrosirius red e Oil Red O, na região anterior da L1, o início do desenvolvimento de hastes quitinosas, a porção inicial do intestino e o primórdio genital. Na região anterior de L2, papilas, bastões quitinosos justapostos à boca e ao intestino maior que em L1. A musculatura de L3 é bem desenvolvida, próximo às hastes quitinosas, há duas estruturas redondas dispostas distalmente uma da outra. Observaram-se também toda a extensão do intestino, o primórdio genital e a intensa celularidade na porção distal da L3. Com o picrosirius red observou-se a musculatura de L1, L2 e L3, assim como, gânglios nervosos na L3. Oil Red O revelou que L1, L2 e L3 armazenam energia em gotículas lipídicas.(AU)

Bovine genital campylobacteriosis: main features and perspectives for diagnosis and control / Campilobacteriose genital bovina: principais características e perspectivas para o diagnóstico e controle

Bovine genital campylobacteriosis (BGC) is a venereal disease caused by Campylobacter fetus subsp. venerealis. In countries with large cattle herds, such as Brazil, where the use of natural breeding as a reproductive strategy is a common practice, BGC is considered an important cause of reproductive failure and economic losses. In these cases, the bull is the asymptomatic carrier of the bacterium and the infected females can have infertility and even abortions. The techniques for the diagnosis of C. fetus are isolation in culture medium and identification by biochemical tests, immunofluorescence, immunoenzymatic assays and molecular techniques. Disease control is based on vaccination with bacterins. This review described the epidemiology, etiology, pathogenesis, and advances in the diagnosis and control of BGC.(AU)

A campilobacteriose genital bovina (CGB) é uma importante enfermidade de caráter venéreo causada por Campylobacter fetus subsp. venerealis. Em países com grandes rebanhos bovinos, como o Brasil, onde o uso da monta natural como estratégia reprodutiva é uma prática corrente, a CGB é considerada uma importante causa de falhas reprodutivas e perdas econômicas. Nestes casos, o touro é o portador assintomático da bactéria e as fêmeas infectadas podem apresentar infertilidade e até mesmo abortos. As técnicas para o diagnóstico de C. fetus são o isolamento em meio de cultura e identificação por testes bioquímicos; imunofluorescência; ensaios imunoenzimáticos e técnicas moleculares. O controle da doença é baseado em vacinação. Neste sentido, esta revisão consiste em uma abordagem sobre a epidemiologia, a etiologia, a patogenia, os avanços no diagnóstico e controle da CGB.(AU)

Papiloma peniano associado a parafimose persistente em um equino / Penile papillomatosis associated with persistent paraphimosis in a horse

Background: Papillomas are cutaneous neoplasms, also known as warts. They are usually benign and are caused by apapillomavirus. The development of papillomas in certain locations on the body may cause irreparable consequences.Paraphimosis is a urological emergency characterized by the inability of the penis to retract or the impossibility of retentioninside the foreskin, causing local circulatory disorders and severe pain. However, the association between genital papillomas and the development of paraphimosis in horses has not been previously documented. The objective here is to describethe clinical and histopathological aspects of a case of penile papilloma associated with persistent paraphimosis in a horse.Case: A 15-year-old mixed-breed, 350 kg, horse presented nodular and crusted lesions, similar to warts, on the penis andforeskin, which progressed over at least 6 months. An incisional biopsy of one of the nodular lesions of the horses peniswas performed. Tissue fragments were collected, packed in 10% buffered formaldehyde, and sent for histopathologicalevaluation to the Animal Pathology Laboratory of the University Veterinary Hospital of the Federal University of CampinaGrande (UFCG), Campus de Patos, Paraíba. The biopsy resulted in a histopathological diagnosis of papilloma, and thehorse was reevaluated. Due to the severity of the clinical case, it was referred to the HVU/UFCG Large Animal Medicaland Surgical Clinic for surgical removal of the penis. The penectomy product was sent to the Animal Pathology Laboratory. Macroscopically, the penis fragment measured 18.0×10.5×6.0 cm in size, had an irregular surface, and presented withnumerous multilobulated, reddish nodules on a sessile base, which were exophytic with projections having the appearanceof a “cauliflower.” The nodules extended from the foreskin and compromised from...(AU)