Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients

Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at ­80o C for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at ­20o C for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at ­80o C for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at ­20o C.(AU)

Enzyme-linked immunosorbent assay (ELISA) for detection of IgY Anti-Borrelia anserina antibodies in Gallus gallus domesticus / Ensaio de imunoadsorção enzimático (ELISA) indireto para detecção de anticorpos IgY Anti-Borreliaanserina em Gallus gallus domesticus

This study aimed to promote the standardization of an indirect, enzyme-linked immunosorbent assay (ELISA) for the serological detection of B. anserina in Gallus gallusdomesticus. An aliquoted sera from vaccinated chicken with B. anserina antigen (GI), experimental infected chickens with B. anserina (GII) and rustic poultry rearing of G. gallus (GIII) were tested with in-house ELISA developed to detect serum antibodies against B. anserina in G. gallus domesticus. On average, the experimentally infected chickens became positive at 9 DPI a mean ± standard deviation (SD) ODI value of 163.11 ± 70.65. The highest observed Optical Density Index (ODI) was 372.54 ± 132.39, at 26 DPI, and the highest overall ODI value was 626.51. The vaccinated chickens became positive between 8 and 10 DPV, with an ODI of 245.59 at 10 DPV, with an overall maximum ODI of 543.13. A total of 108 blood samples were collected from poultry raised on rustic farms. Of the total samples collected, 58.33% (63/108) were considered positive for B. anserina. The maximum ODI found among these rustic chickens was 283.24. This stardardization provided a sensitivity and specificity of 100%.

Este estudo teve como objetivo promover a padronização de um ensaio imunoenzimático indireto (ELISA) para a detecção sorológica de Borrelia anserina em Gallus gallus domesticus. Um frango vacinado com antígeno de B. anserina (GI), frangos infectados experimentalmente com B. anserina (GII) e frangos criados de forma rústica (GIII) foram testados com ELISA indireto in house desenvolvido para a detecção sorológica contra B. anserina em G. gallus domesticus. Em média, os frangos infectados experimentalmente tornaram-se positivos aos 9º dia pós-inoculação (DPI), um valor do índice de densidade óptica (ODI) médio ± desvio padrão (SD) de 163,11 ± 70,65. O maior ODI observado foi 372,54 ± 132,39, em 26ºDPI, e o maior valor geral de ODI foi 626,51. Os frangos vacinados tornaram-se positivos entre 8º e 10° DPV, com um ODI de 245,59 a 10 DPV, com um ODI máximo geral de 543,13. Um total de 108 amostras de sangue foram coletadas de aves criadas em fazendas rústicas. Do total de amostras coletadas, 58,33% (63/108) foram consideradas positivas para B. anserina. O ODI máximo encontrado entre essas galinhas rústicas foi 283,24. Essa padronização proporcionou sensibilidade e especificidade de 100%.

Canine visceral leishmaniasis in Rio Grande do Norte state, Northeastern Brazil - spatial analysis

Background: Human visceral leishmaniasis (HVL) is a potentially fatal disease with a worldwide distribution, being endemic in 12 countries in the Americas. The main reservoir in the urban environment is the dog, whose cases precede the disease in humans. For the control of HVL, the Ministry of Health of Brazil recommends efficiency in the notification of human cases, control of sandflies, elimination of reservoirs and health education, in addition to the interruption in the transmission of the disease by the intensification of surveillance and control of priority areas based on identification by spatial analysis. The objective of the study was to investigate the spatial distribution of canine visceral leishmaniasis (CVL) in the state of Rio Grande do Norte, Brazil, determining areas of risk by identifying spatial clusters, with a view to monitoring and implementing preventive actions. Materials, Methods & Results: Secondary data from sample and/or routine serological surveys for serological diagnosis of LVC in the period from 2011 to 2018 were used. The inclusion of animals in the routine diagnosis per municipality resulted from demands of veterinarians, veterinary clinics, dog tutors, zoonoses control centers and environmental surveillance. The spatial statistical analysis was performed with SatScan software version 9.6 for the detection of spatial clusters, based on using the statistical scan method. Of the total of 231,123 dogs tested in the period, 24,642 (10.6%) were seroreactive for CVL. During the study, the municipalities with the highest number of cases were Natal and Mossoró, with 9,671 and 4,514 cases, respectively. During the years 2011 to 2018, 38 significant clusters (P < 0.05) were identified that included one or more municipalities. Discussion: The state of Rio Grande do Norte has an urban environment susceptible to the occurrence of CVL, with climate and topography that favor the proliferation of the vector and housing in precarious socio-sanitary conditions. The high number of CVL cases in Natal can be explained by the fact that the city is considered endemic for CVL, characterized as an area of intense transmission of the disease, according to the Ministry of Health. In addition, public infrastructure in some locations is deficient and living conditions are unfavorable, so that there is a need to invest in effective protection measures for vector control, as well as a focus on health education, whose HVL control measures in the municipality need to be readjusted. The high rate of cases and the constant presence of clusters in the municipality of Açu can be explained by the increasing degradation of the Caatinga biome, evidenced by the removal of firewood for use in the ceramist pole, whose activity is concentrated on a large scale in the use of raw material and energy, through the production of charcoal, for agricultural and livestock fronts, putting species of fauna and flora at risk. It is also noteworthy that this fact contributes to the destruction of wild ecotopes, resulting in the search for the vector for other sources of human and animal food, allowing an increase in the number of cases of the disease. It's concluded that canine visceral leishmaniasis is distributed in a large part of the state of Rio Grande do Norte. The underreporting and/or deficiency in the disclosure of data by some municipalities represents a challenge in complying with the actions of the Visceral Leishmaniasis Surveillance and Control Program, and attention should be paid to the monitoring and inspection of the execution actions of municipal managers, as well as how to train professionals who are part of the service.

Blue tongue virus in dairy cattle in the Southern region of Minas Gerais, Brazil - serological survey

Background: Blue tongue (BT) is a noncontagious viral disease transmitted by hematophagous arthropods, especially of the genus Culicoides. The economic impact of the disease is related not only to deaths in sheep herds but also to the possible correlation of virus infection with the development of other diseases, such as pneumonia, abortion and movement problems. The economic losses caused by Blue Tongue are linked to restrictions on the import and export of animals and their genetic material and to the reproductive disorders associated with this disease. In addition, the fact that cattle take the role of reservoir, combined with the care by other countries with outbreaks of infection and biological contamination of their products, hinders trade in Mercosul, United States and Europe. Cattle are affected by Blue Tongue Virus in endemic areas and in some epidemic areas, but the development of clinical disease is rare. The clinical signs, when evident, range from reproductive losses, such as embryonic death, abortion, fetal malformation, temporary sterility, infertility in bulls, stillbirths and the birth of weak animals. The objective of this study was to determine the epidemiological aspects of Blue Tongue Virus (BTV) infection in dairy cattle in the Lavras region, state of Minas Gerais, Brazil. Materials, Methods & Results: A cross-sectional study was conducted to evaluate the frequency of cattle and herds seropositive for Blue Tongue in the southern region of Minas Gerais. In this study, 54 dairy farms were visited. A total of 586 serum samples were collected from cows of reproductive age. Sampling was random, and serum samples were collected from lactating cows over 24 months of age by puncture of the jugular vein and/or coccidian vein. The samples were transported and stored at the Setor de Patologia Veterinária, at the Universidade Federal de Lavras (SPV-UFLA), where they were centrifuged, and the serum aliquots were obtained, transferred to microtubes and kept at -20°C until the serological tests were performed. The samples were tested with the agarose gel immunodiffusion test (AGID) for anti-blue tongue virus antibodies. The AGID test is more practical and is the main method used to identify Blue Tongue Virus seroprevalence in different ruminant species. They are considered important tools for epidemiological surveillance of the disease. A prevalence of 83.28% was observed among animals that were seropositive for Blue Tongue Virus (488/586; IC 95% = 80.0 - 86.21). In addition, 100% (54/54; IC 95% = 93.4 - 100.0) of the farms had at least 1 positive animal, with rates ranging from 45.45% to 100% within the herds and where 22.22% of the farms had rates of 100% of the animals being positive. Discussion: Blue Tongue is a disease known to affect domestic and wild ruminants in Brazil. However, there is a lack of more precise information about its epidemiology and occurrence in the country and of joint efforts of researchers, producers and the government to understand in detail both the biology of vectors and the viral biology of Blue Tongue Virus in Brazil. This is the first record of detection of anti-blue tongue virus antibodies in cattle in the southern region of Minas Gerais. The results suggest that Blue Tongue Virus is present in cattle in the study area.

Ocorrência de Babesia sp, Ehrlichia canis e Hepatozoon canis em cães domiciliados, em dois municípios do estado do Espírito Santo - Brasil / Occurrence of Babesia sp, Ehrlichia canis and Hepatozoon canis in domiciled dogs in two municipalities of the state of Espírito Santo - Brazil / Ocurrencia de Babesia sp, Ehrlichia canis y Hepatozoon canis em perros domiciliados em dos municipios del estado de Espírito Santo - Brasil

As doenças transmitidas por carrapatos são afecções de grande importância na clínica médica de pequenos animais, devido à alta casuística e ampla distribuição vetorial no território brasileiro. Os principais agentes responsáveis pelas infecções em cães são Babesia sp., Ehrlichia canis e Hepatozoon canis. Os animais infectados são assintomáticos ou apresentam sinais clínicos inespecíficos, sendo necessário a utilização de testes diagnósticos para definição do agente etiológico, e diagnóstico seguro. O objetivo do presente estudo foi determinar a ocorrência desses micro-organismos em cães naturalmente infectados, domiciliados nos municípios de Vila Velha e Anchieta, Espírito Santo, utilizando diferentes testes de detecção: Reação em cadeia polimerase (PCR), sorologia para detecção de anticorpos anti Ehrlichia canis e pesquisa de hematozoários em esfregaço sanguíneo. Foram analisadas 65 amostras de sangue obtidas por venopunção de veia cefálica de cães. No teste de PCR, 4,62% dos animais foram positivos para Babesia vogeli e 1,54% para Ehrlichia canis sendo os resultados para Hepatozoon canis negativos. No teste sorológico para E. canis 90,77% dos animais foram positivos para a presença de anticorpos, e na pesquisa em lâminas de esfregaço sanguíneo 3,02% apresentavam outros hemoparasitas. Os resultados indicam a dispersão desses hemoparasitas na população canina da região de estudo, entretanto com baixa ocorrência. O teste de PCR demonstrou-se como o mais sensível no qual Babesia vogeli foi o agente mais observado.(AU)

Tick-borne diseases are diseases of great importance in the medical practice of small animals, due to the high casuistry and wide vectorial distribution in the Brazilian territory. The main agents responsible for infections in dogs are Babesia sp., Ehrlichia canis and Hepatozoon canis. Infected animals are asymptomatic or present nonspecific clinical signs, requiring the use of diagnostic tests to define the etiologic agent, and safe diagnosis. The objective of the present study was to determine the occurrence of these microorganisms in naturally infected dogs domiciled in the municipalities of Vila Velha and Anchieta, Espírito Santo, using different detection tests: polymerase chain reaction (PCR), serology to detect antibodies against Ehrlichia canis and research of hematozoa in blood smears. Sixty-five blood samples obtained by venipuncture of the cephalic vein of dogs were analyzed. In the PCR test, 4.62% of the animals were positive for Babesia vogeli and 1.54% for Ehrlichia canis, and the results for Hepatozoon canis were negative. In the serological test for E. canis, 90.77% of the animals were positive for the presence of antibodies, and in the research in blood smear slides, 3.02% presented other hemoparasites. The results indicate the dispersion of these hemoparasites in the canine population of the study region, however with low occurrence. The PCR test proved to be the most sensitive, in which Babesia vogeli was the most observed agent.(AU)

Las enfermedades transmitidas por garrapatas son enfermedades de gran importancia en la práctica médica de los pequeños animales, debido a la alta casuística y amplia distribución vectorial en el territorio brasileño. Los principales agentes responsables de las infecciones en los perros son Babesia sp., Ehrlichia canis y Hepatozoon canis. Los animales infectados son asintomáticos o presentan signos clínicos inespecíficos, siendo necesario el uso de pruebas diagnósticas para la definición del agente etiológico, y el diagnóstico seguro. El objetivo del presente estudio fue determinar la ocurrencia de estos microorganismos en perros infectados naturalmente, domiciliados en los municipios de Vila Velha y Anchieta, Espírito Santo, utilizando diferentes pruebas de detección: reacción en cadena de la polimerasa (PCR), serología para detectar anticuerpos anti Ehrlichia canis e investigación de hematozoos en frotis de sangre. Se analizaron sesenta y cinco muestras de sangre obtenidas por venopunción de la vena cefálica de los perros. En la prueba PCR, el 4,62% de los animales fueron positivos para Babesia vogeli y el 1,54% para Ehrlichia canis, y los resultados para Hepatozoon canis fueron negativos. En la prueba serológica para E. canis, el 90,77% de los animales fueron positivos a la presencia de anticuerpos, y en la investigación en láminas de frotis de sangre el 3,02% presentaron otros hemoparásitos. Los resultados indican la dispersión de estos hemoparásitos en la población canina de la región de estudio, aunque con una baja presencia. La prueba PCR resultó ser la más sensible, en la que Babesia vogeli fue el agente más observado.(AU)

Occurrence of Leptospira spp. and factors associated with the infection in horses from a military contingent in the Rio Grande do Sul State, Brazil / Ocorrência de Leptospira spp. e fatores associados à infecção em equinos de contingente militar no estado do Rio Grande do Sul, Brasil

The seroprevalence of Leptospira spp. and infection risk factors were investigated in horses from a military contingent in the Rio Grande do Sul State, Brazil. A total of 446 horses were evaluated and categorized into 309 mares, 11 stallions, and 126 horses with an average age of three years. To determine seroprevalence, serum samples from all horses were submitted to the microscopic agglutination test against 12 serovars belonging to nine serogroups, usually circulating in equine populations. To investigate the possible risk factors of infection by Leptospira spp., questionnaires were applied in that military unit. The seroprevalence in the horses with an average age of 3 years, mares, and stallions were 57.94% (73/126), 54.05% (165/309), and 45.45% (5/11), respectively. The main risk factors identified were contact with different water sources, the presence of rodents, and contact with wild and domestic animals. The high seroprevalence of anti­Leptospira spp. antibodies may be associated with the presence of these risk factors. Therefore, the exposure of horses to possible reservoirs of Leptospira spp. should be minimized. Also, the immunoprophylaxis protocol should be reviewed and a shorter interval between vaccinations adopted to control leptospirosis in this herd.(AU)

A soroprevalência de Leptospira spp. e os fatores de risco para infecção foram investigados em cavalos de um contingente militar no Rio Grande do Sul, Brasil. Um total de 446 cavalos foram avaliados e categorizados em 309 éguas, 11 garanhões e 126 cavalos com idade média de três anos. Para determinação da soroprevalência, amostras de soro de todos os equinos foram submetidas ao teste de aglutinação microscópica contra 12 sorovares pertencentes a nove sorogrupos, normalmente circulantes em populações equinas. Para investigar os possíveis fatores de risco da infecção por Leptospira spp., foi aplicado um questionário naquela unidade militar. A soroprevalência nos cavalos com idade média de três anos, éguas e garanhões foi de 57,94% (73/126), 54,05% (165/309) e 45,45% (5/11), respectivamente. Os principais fatores de risco identificados foram o contato com diferentes fontes de água, a presença de roedores e o contato com animais silvestres e domésticos. A alta soroprevalência de anticorpos anti­Leptospira spp. pode estar associada à presença desses fatores de risco. Portanto, a exposição destes equinos aos possíveis reservatórios de Leptospira spp. deve ser minimizada. Além disso, o protocolo de imunoprofilaxia deve ser revisto e, possivelmente, um menor intervalo entre as vacinas deve ser adotado para o controle da leptospirose neste plantel.(AU)

Western Blot no imunodiagnóstico de lentivírus de pequenos ruminantes / Western Blot in immunodiagnosis of small ruminant lentivirus

Background: Small ruminant lentivirus (SRLV) belong to genus Lentivirus, family Retroviridae. These viruses causecaprine arthritis encephalitis (CAE) and maedi visna (MV), infectious diseases that cause economic, production, and reproductive losses. There are no effective treatments or vaccines for these diseases. Thus, early detection via serology hasgreat importance for control of SRLV. Therefore, the objective of this review is to demonstrate the potential of the westernblot (WB) test as an immunodiagnostic test for SRLV.Review: In general, immunodiagnosis of SRLV is performed via agar gel immunodiffusion (AGID) and indirect enzymelinked immunosorbent assay (ELISA), which can detect antibodies in several different biological samples but is used preferably with serum and blood plasma. However, WB has demonstrated efficacy in the early diagnosis of immunoglobulinsagainst SRLV, presenting higher sensitivity and specificity than the serological tests usually used, because this techniquecan detect antibodies at a dilution as much as 256 times greater than that of AGID and 32 times greater than that of ELISA.SRLV infection and consequent immunological activation result in the induction of cellular and humoral responses. Additionally, around the third week, production of antibodies directed mainly toward viral capsid proteins (p25 and p28)occurs. After the fifth week, production of immunoglobulins directed toward other viral proteins occurs. Because of thepersistence of SRLV infection, serology is considered to be the most practical means to diagnosis. Each serological testhas a percentage specificity and distinct sensitivity, as well as advantages and disadvantages in its applicability. It shouldbe noted that there is no gold standard test for diagnosis of SRLV infection. Moreover, SRLV are characterized by escapemechanisms such as genetic diversity, mutagenic potential, viral intermittence...

Occurrence of Leptospira spp. and factors associated with the infection in horses from a military contingent in the Rio Grande do Sul State, Brazil / Ocorrência de Leptospira spp. e fatores associados à infecção em equinos de contingente militar no estado do Rio Grande do Sul, Brasil

The seroprevalence of Leptospira spp. and infection risk factors were investigated in horses from a military contingent in the Rio Grande do Sul State, Brazil. A total of 446 horses were evaluated and categorized into 309 mares, 11 stallions, and 126 horses with an average age of three years. To determine seroprevalence, serum samples from all horses were submitted to the microscopic agglutination test against 12 serovars belonging to nine serogroups, usually circulating in equine populations. To investigate the possible risk factors of infection by Leptospira spp., questionnaires were applied in that military unit. The seroprevalence in the horses with an average age of 3 years, mares, and stallions were 57.94% (73/126), 54.05% (165/309), and 45.45% (5/11), respectively. The main risk factors identified were contact with different water sources, the presence of rodents, and contact with wild and domestic animals. The high seroprevalence of anti­Leptospira spp. antibodies may be associated with the presence of these risk factors. Therefore, the exposure of horses to possible reservoirs of Leptospira spp. should be minimized. Also, the immunoprophylaxis protocol should be reviewed and a shorter interval between vaccinations adopted to control leptospirosis in this herd.(AU)

A soroprevalência de Leptospira spp. e os fatores de risco para infecção foram investigados em cavalos de um contingente militar no Rio Grande do Sul, Brasil. Um total de 446 cavalos foram avaliados e categorizados em 309 éguas, 11 garanhões e 126 cavalos com idade média de três anos. Para determinação da soroprevalência, amostras de soro de todos os equinos foram submetidas ao teste de aglutinação microscópica contra 12 sorovares pertencentes a nove sorogrupos, normalmente circulantes em populações equinas. Para investigar os possíveis fatores de risco da infecção por Leptospira spp., foi aplicado um questionário naquela unidade militar. A soroprevalência nos cavalos com idade média de três anos, éguas e garanhões foi de 57,94% (73/126), 54,05% (165/309) e 45,45% (5/11), respectivamente. Os principais fatores de risco identificados foram o contato com diferentes fontes de água, a presença de roedores e o contato com animais silvestres e domésticos. A alta soroprevalência de anticorpos anti­Leptospira spp. pode estar associada à presença desses fatores de risco. Portanto, a exposição destes equinos aos possíveis reservatórios de Leptospira spp. deve ser minimizada. Além disso, o protocolo de imunoprofilaxia deve ser revisto e, possivelmente, um menor intervalo entre as vacinas deve ser adotado para o controle da leptospirose neste plantel.(AU)

Epidemiological study of dengue fever in district Swabi, Khyber Pakhtunkhwa, Pakistan / Estudo epidemiológico da dengue no distrito Swabi, Khyber Pakhtunkhwa, Paquistão

Dengue is a viral disease which is serious health concern from last few decades and the infection transmitted through mosquito bite into human. This study was conducted to carry out prevalence of dengue fever in District Swabi. A total of 196 blood sample were collected from patients with age ranges (0-80 years) having dengue fever on the basis of physical symptoms from Bacha Khan Medical Complex Swabi during August to October 2017. Serological test were performed for detection of IgM, IgG and NS1 (Non structural protein antigen of virus) against dengue. Out of total 196 confirmed dengue cases the most prone gender was male 123(62%) while 73(38%) were female. Among the age groups; 21-30 years group 62 (31.6%) was found the most predominated age group. The higly affected areas in district Swabi were Topi (40.8%) followed by Main Swabi (27%), Maneri (8.2%), Marghuz (6%), Shawa Ada (5.1%), Shah Mansoor (5.1%), Gohati (4.1%), and Chota Lahore (3.6%). Therefore, the health department should take actions by educating the public about basic cleanliness of the environment. The community should be encouraged to participate in the control of such vector based diseases/infections.(AU)

A dengue é uma doença viral que é um sério problema de saúde das últimas décadas e a infecção transmitida através da picada de mosquito em humanos. Este estudo foi realizado para realizar a prevalência da dengue no distrito de Swabi. Foram coletadas 196 amostras de sangue de pacientes com faixa etária (0 a 80 anos) com dengue com base nos sintomas físicos do Complexo Médico Bacha Khan de Swabi, no período de agosto a outubro de 2017. Foram realizados testes sorológicos para detecção de IgM, IgG e NS1 (antígeno proteico não estrutural do vírus) contra a dengue. Do total de 196 casos confirmados de dengue, o sexo mais propenso foi o masculino 123 (62%), enquanto 73 (38%) eram do sexo feminino. Entre as faixas etárias; A faixa etária de 21 a 30 anos 62 (31,6%) foi a faixa etária mais predominante. As áreas altamente afetadas no distrito de Swabi foram Topi (40,8%), seguidas por Main Swabi (27%), Maneri (8,2%), Marghuz (6%), Shawa Ada (5,1%), Shah Mansoor (5,1%), Gohati (4,1%) e Chota Lahore (3,6%). Portanto, o departamento de saúde deve tomar ações educando o público sobre a limpeza básica do meio ambiente. A comunidade deve ser incentivada a participar do controle de tais doenças / infecções baseadas em vetores.(AU)

Effect of Essential Oil and Organic Acid on Performance, Gut Health, Bacterial Count and Serological Parameters in Broiler

A total of 1500 day old broiler chicks were grouped into five treatments; each treatment group further consisted of six replicates. Group A was given a control diet having antibiotics. Group B and C were offered essential oil blend at the proportion of 0.1 g/kg and organic acid at the dose rate of 1g/kg. Group D was given both essential oil 0.1 g/kg and organic acid 1g/kg in combination. Group E was offered only a basal diet without antibiotics and considered as a negative control. The trial lasted for 35 days. Results indicated that the body weight gain and feed conversion ratio (FCR) exhibited significant improvement but insignificant in the case of feed consumption. Bodyweight and FCR were better in group B, followed by C. Carcass characteristics like eviscerated weight and giblet weight were also improved in group B while dressed weight showed insignificant results. Total cholesterol and high-density lipoproteins (HDL) were unaffected by the treatments; however, increased blood glucose levels and decreased low-density lipoproteins (LDL) concentration were significantly noticed. Treatment groups have no effect on antibody titer against ND at the end of the trial. In gut morphology, significantly higher villus height was observed in group C, but villus width and crypt depth remained unaffected. In conclusion, essential oil and organic acids have improved performance, carcass parameters, serum biochemistry, gut health, and decreased bacterial count.(AU)

Western Blot no imunodiagnóstico de lentivírus de pequenos ruminantes / Western Blot in immunodiagnosis of small ruminant lentivirus

Background: Small ruminant lentivirus (SRLV) belong to genus Lentivirus, family Retroviridae. These viruses causecaprine arthritis encephalitis (CAE) and maedi visna (MV), infectious diseases that cause economic, production, and reproductive losses. There are no effective treatments or vaccines for these diseases. Thus, early detection via serology hasgreat importance for control of SRLV. Therefore, the objective of this review is to demonstrate the potential of the westernblot (WB) test as an immunodiagnostic test for SRLV.Review: In general, immunodiagnosis of SRLV is performed via agar gel immunodiffusion (AGID) and indirect enzymelinked immunosorbent assay (ELISA), which can detect antibodies in several different biological samples but is used preferably with serum and blood plasma. However, WB has demonstrated efficacy in the early diagnosis of immunoglobulinsagainst SRLV, presenting higher sensitivity and specificity than the serological tests usually used, because this techniquecan detect antibodies at a dilution as much as 256 times greater than that of AGID and 32 times greater than that of ELISA.SRLV infection and consequent immunological activation result in the induction of cellular and humoral responses. Additionally, around the third week, production of antibodies directed mainly toward viral capsid proteins (p25 and p28)occurs. After the fifth week, production of immunoglobulins directed toward other viral proteins occurs. Because of thepersistence of SRLV infection, serology is considered to be the most practical means to diagnosis. Each serological testhas a percentage specificity and distinct sensitivity, as well as advantages and disadvantages in its applicability. It shouldbe noted that there is no gold standard test for diagnosis of SRLV infection. Moreover, SRLV are characterized by escapemechanisms such as genetic diversity, mutagenic potential, viral intermittence...(AU)

Effect of Essential Oil and Organic Acid on Performance, Gut Health, Bacterial Count and Serological Parameters in Broiler

A total of 1500 day old broiler chicks were grouped into five treatments; each treatment group further consisted of six replicates. Group A was given a control diet having antibiotics. Group B and C were offered essential oil blend at the proportion of 0.1 g/kg and organic acid at the dose rate of 1g/kg. Group D was given both essential oil 0.1 g/kg and organic acid 1g/kg in combination. Group E was offered only a basal diet without antibiotics and considered as a negative control. The trial lasted for 35 days. Results indicated that the body weight gain and feed conversion ratio (FCR) exhibited significant improvement but insignificant in the case of feed consumption. Bodyweight and FCR were better in group B, followed by C. Carcass characteristics like eviscerated weight and giblet weight were also improved in group B while dressed weight showed insignificant results. Total cholesterol and high-density lipoproteins (HDL) were unaffected by the treatments; however, increased blood glucose levels and decreased low-density lipoproteins (LDL) concentration were significantly noticed. Treatment groups have no effect on antibody titer against ND at the end of the trial. In gut morphology, significantly higher villus height was observed in group C, but villus width and crypt depth remained unaffected. In conclusion, essential oil and organic acids have improved performance, carcass parameters, serum biochemistry, gut health, and decreased bacterial count.

Comparison of serological and molecular tests to investigate Leishmania spp. infections in stray dogs from an area of intense visceral leishmaniasis transmission in Brazil / Comparação de testes sorológicos e moleculares para investigar infecções por Leishmania spp. em cães errantes de uma área de intensa transmissão de leishmaniose visceral no Brasil

The aim of this study was to investigate the level of exposure to Leishmania infection in stray dogs in an area of intense visceral leishmaniasis transmission in the state of Pernambuco, Brazil. Blood samples from 178 dogs were analyzed using serological and molecular assays: rapid immunochromatographic test (ICT), enzyme-linked immunosorbent assay (ELISA), immunofluorescence antibody test (IFAT), and conventional and quantitative polymerase chain reaction (cPCR and qPCR). Positivity values obtained with serological tests were 71.4% (127/178), 70.2% (125/178), and 50.6% (90/178) using ICT, ELISA, and IFAT, respectively, with 38.8% (69/178) of the dogs were simultaneously positive for all three tests. The positivity values obtained with cPCR and qPCR were 20.2% (36/178) and 38.8% (69/178), respectively, with 11.8% (21/178) testing positive in both molecular assays. Overall, 87.1% (155/178) were positive for anti-Leishmania spp. antibodies and/or Leishmania spp. DNA. Positivity to one or more tests was statistically associated with lymphadenomegaly, skin lesions, lymphocytosis, anemia and hyperproteinemia. The results of this study revealed a high level of exposure to Leishmania in stray dogs in an area of intense human visceral leishmaniasis transmission, suggesting that dogs play a role as reservoirs in the transmission cycle of this zoonosis.(AU)

O objetivo deste estudo foi investigar o nível de exposição à infecção por Leishmania em cães errantes de uma área de transmissão intensa de leishmaniose visceral no estado de Pernambuco. Amostras de sangue de 178 cães foram avaliadas por testes sorológicos e moleculares: teste imunocromatográfico (IC), ensaio imunoenzimático (ELISA), teste de imunofluorescência de anticorpos (RIFI), reação em cadeia da polimerase convencional e quantitativa (cPCR e qPCR). Os valores de positividade obtidos com os testes sorológicos foram 71,4% (127/178), 70,2% (125/178) e 50,6% (90/178) com IC, ELISA e RIFI, respectivamente. E com 38,8% (69/178) dos cães mostraram-se simultaneamente positivos aos três testes. Os valores de positividade obtidos por cPCR e qPCR foram 20,2% (36/178) e 38,8% (69/178), respectivamente. E com 11,8% (21/178) dos cães foram positivos em ambos os testes moleculares. No geral, 87,1% (155/178) foram positivos para anticorpos anti-Leishmania spp. e/ou material genético de Leishmania spp. A positividade a um ou mais testes foi estatisticamente associada à linfonodomegalia, lesões cutâneas, linfocitose, anemia e hiperproteinemia. Os resultados do estudo revelam um alto nível de exposição à Leishmania spp. em cães errantes em uma área de transmissão intensa de leishmaniose visceral humana, sugerindo que eles podem desempenhar um papel como reservatórios no ciclo de transmissão dessa zoonose.(AU)

Leishmaniose visceral em cães / Visceral leishmaniasis in dogs

Background: Canine visceral leishmaniasis (CVL) is a zoonosis of variable clinical presentation, either in systemic orcutaneous form. Clinical signs include anorexia, ophthalmopathies, and chronic kidney disease. In the state of Santa Catarina, the foci are concentrated in the capital and its adjacencies, in the east side of the state. The objective of this studyis to outline the first three reported cases of CVL in the municipality of Curitibanos, since there are no reports to date inthe region of the mountainous plateau, in the middle west of Santa Catarina.Cases: All dogs were treated at the Veterinary Clinic School of the Federal University of Santa Catarina, Campus Curitibanos. The animals, two males and one female, belonged to the same tutor, resided in Curitibanos, and were attendedbetween 2016 and 2020. In the first case, attended in 2016, the complaint was of eye and skin changes about three monthsago. The animal lived in an urban environment and came from Uruguaiana, Rio Grande do Sul. On physical examination,the animal presented skin peeling, wet and crusty lesions, bloody discharge in the ears and nasal hyperkeratosis, as wellas signs suggestive of uveitis. In this case, euthanasia was carried out. The second case had complaints of respiratory, eyechanges, hyporexia, and polydipsia. The dog was prostrate, dehydrated, with lymphadenomegaly and respiratory disorders, compatible with bacterial pneumonia. Bilateral corneal edema confirmed uveitis. Upon return, the animal remaineddehydrated with enlarged lymph nodes. In the aspiration of the lymph nodes, suggestive forms of Leishmania sp. wereobserved. The recommended confirmatory tests were performed, leading to a definitive diagnosis of CVL. The patientwas treated with miltefosine, but later died. The third case was attended for general evaluation after a positive diagnosisfor CVL during an epidemiological survey of the second case. The animal was alert, tachycardic, and tachypneic with...(AU)

Serologic profile of bovine Alfaherpesvirus type1 (BoHV-1) and Pestivírus A (BVDV-1) in herds of a family farming of the Sao Paulo state / Perfil sorológico do Alfaherpesvirus bovino tipo 1 (BoHV-1) e Pestivírus A (BVDV-1) em rebanhos da agricultura familiar no estado de São Paulo

Serum samples from 719 dairy cattle of reproductive age, from 47 farms in a settlement in the municipality of Andradina, São Paulo, were analyzed for the presence of antibodies against the Nebraska strain of BoHV-1 and Singer type 1 of BVDV-1, using the virus neutralization technique (VN). Regarding BoHV-1, 72.0% of the samples analyzed (518/719) were positive, with a geometric mean of antibody titers of 318. For BVDV-1, 44.8% of the samples were reactive, only one farm had no reactive animals, and the majority of antibody titers found in the animals' sera were low. The results of this study revealed that BoHV-1 was present in all of the studied herds, with moderate titration in the majority of cases, with the presence of some high individual titers among the tested animals. Also, even with the reactivation or circulation of BoHV-1 in herds, the clinical disease was not observed, an epidemiological finding that also applied to BVDV-1. In view of the results obtained, the circulation of diseases in family farm herds in the studied region was evident, suggesting the need to study local risk factors and improve health education policies as a way to prevent or reduce the prevalence of these diseases.

Coletou-se 719 amostras de soro sanguíneo de bovinos em idade reprodutiva, de 47 propriedades de um assentamento no município de Andradina-SP. As amostras foram analisadas para presença de anticorpos contra ambos os vírus estudados, pela técnica de virusneutralização, utilizando-se a estirpe Nebraska para o BoHV-1 e Singer tipo 1 para o BVDV-1. Em relação ao BoHV-1, 72,0% (518/719) das amostras foram positivas, sendo 318 a média geométrica dos títulos de anticorpos (GMT).Para o BVDV-1, 44,8% das amostras foram reagentes; apenas uma propriedade não teve animais reagentes, sendo a maioria dos títulos de anticorpos encontrados nos soros baixa. Observou-se que o BoHV-1 está presente em todos os rebanhos estudados com titulação moderada, com presença de alguns títulos individuais altos entre animais e com GMT alta em um dos rebanhos. Mesmo havendo a reativação ou mesmo a circulação do BoHV-1 nos rebanhos, não foi observado a doença clínica, fato epidemiológico que também ocorreu com o BVDV-1. Diante dos resultados obtidos, ficou evidente a circulação das doenças nos rebanhos da agricultura familiar na região estudada, sugerindo o estudo de fatores de risco local e melhorias de políticas de educação em saúde como forma de prevenir e ou reduzir a prevalência das enfermidades.

Serological survey of leptospirosis, brucellosis, and lentivirus in herds of small ruminants in Recôncavo Baiano, Bahia, Brazil / Inquérito sorológico de leptospirose, brucelose e lentiviroses em rebanhos de pequenos ruminantes no Recôncavo Baiano, Bahia, Brasil

A serological survey was carried out to assess the frequency of leptospirosis, small ruminants lentivirus (SRLV), and brucellosis in small ruminant herds in the Recôncavo Baiano, State of Bahia, Brazil, from February to December 2017. In four goat herds, 125 animals were tested for SRLV and leptospirosis, while in five sheep herds, 378 animals were tested for leptospirosis, brucellosis, and SRLV. Regarding leptospirosis, MAT detected 80% of goats and 15.34% of sheep seroreactive. Reactivity was most frequent to serogroups Autumnalis and Grippotyphosa in goats and sheep, respectively. Regarding SRLV, 8.8% of goats and 0.79% of sheep were reactive. Search for anti-B. ovis antibodies revealed 0.52% reactivity. In sheep, three animals showed simultaneous seroreactivity for SRLV and leptospirosis, while one animal had a serological response for brucellosis and leptospirosis. In goats, simultaneous seroreactivity for SRLV and leptospirosis was observed in only one animal. Leptospirosis was the most frequent of the three infectious diseases in investigated herds.(AU)

Foi realizado um inquérito sorológico para avaliar a frequência de ocorrência de leptospirose, lentiviroses de pequenos ruminantes (LVPR) e brucelose em rebanhos de pequenos ruminantes no Recôncavo Baiano, estado da Bahia, Brasil, no período de fevereiro a dezembro de 2017. Em quatro rebanhos de caprinos, foram testados 125 animais para LVPR e leptospirose, enquanto em cinco rebanhos de ovinos, foram testados 378 animais para leptospirose, brucelose e LVPR. Em relação à leptospirose, 80% das cabras e 15,34% das ovelhas foram sororreativas. Os sorogrupos de Leptospira spp. predominantes foram Autumnalis e Grippotyphosa para caprinos e ovinos, respectivamente. Em relação as LVPR, 8,8% dos caprinos e 0,79% dos ovinos foram reativos. Adicionalmente, a pesquisa de anticorpos Anti-B. ovis revelou 0,52% de ovinos reativos. Em ovinos, três animais apresentaram sororreatividade simultânea para LVPR e leptospirose, enquanto um animal teve resposta sorológica para brucelose e leptospirose. Em caprinos, sororreatividade simultânea para LVPR e leptospirose foi observada em apenas um animal. A leptospirose foi a doença infecciosa mais frequente nos rebanhos investigados.(AU)

True prevalence and spatial distribution of equine infectious anemia virus (EIAV) in horses from northeast region of Brazil

Equine infectious anemia (EIA) is a viral infection, caused by a lentivirus of the Retroviridae family, Orthoretrovirinael subfamily and its occurrence generates significant economic losses due to culling of positive animals as a measure of infection control. The objective of this work was to determine the prevalence of horses positive for equine infectious anemia virus (EIAV) and to identify the occurrence of areas with higher densities of cases in the states of Paraíba (PB), Pernambuco (PE), Rio Grande do Norte (RN) and Ceará (CE), Northeast region of Brazil, during the rainy (May and June) and dry (October and November) periods of 2017 and 2018. Serum samples from 6,566 horses from the states of PB, PE, RN and CE, Brazil, provided by the Laboratório Veterinária Diagnóstico - Ltda., were used. Serological diagnosis of EIA was performed using indirect enzyme-linked immunosorbent assay (ELISA) as a screening test and agar gel immunodiffusion test (AGID) as a confirmatory test. The apparent prevalence was obtained by dividing the number of seroreactive animals by the total number of animals, while the true prevalence was estimated by adjusting the apparent prevalence, considering the sensitivity (100%) and specificity (98.6%) of the diagnostic protocol used. For the construction of Kernel estimates, the Quartic function was used. In the dry season, of the 1,564 animals sampled, 28 were serologically positive, of which 19 belonged to the state of Ceará, 7 to Paraíba and 2 to Rio Grande do Norte. In 2018, it was observed that, during the rainy season, 26 of the 1,635 horses were seroreactive, with 19 cases resulting from Ceará, 4 from Paraíba and 3 from Pernambuco. In the dry season, 32 of the 1,526 animals were seroreactive to EIAV, of which 26 were from Ceará, 3 from Paraíba, 1 from Rio Grande do Norte and 2 from Pernambuco. In the dry period of 2017, the CE had a real prevalence of 1.22% (95% CI = 0.05 - 2.99%). In 2018, during the rainy season, prevalences of 0.03% (95% CI = 0 - 1.18%) were identified in CE and 1.69% (95% CI = 0 - 8.38%) in PE. Regarding the 2018 dry period, a prevalence of 1.32% (95% CI = 0.26 - 2.84%) was found in the state of CE. In both dry and rainy periods of 2017, the presence of spatial clusters of animals positive for EIA was observed, mainly in the border areas among the states of CE, PE, PB and RN. In 2018, there was a variation in the distribution of areas with higher densities of cases between the rainy and dry periods. The state of CE had the highest prevalence of positive animals and the presence of areas with higher densities of EIA cases in both climatic periods, in the years 2017 and 2018. In some municipalities of the CE, important sporting events of agglomeration of animals take place, which can favor the transmission of EIAV by facilitating the contact of infected and susceptible animals. Population density may be a factor associated with the higher prevalence observed in this region, as it has the second largest herd among the states studied. Higher densities indirectly contribute to the occurrence of infectious diseases, as they favor the contact of infected and susceptible animals. The occurrence of higher densities of cases in the border areas of the states of PE, RN, CE, and PB may be related to the greater movement of animals in these regions, favoring the indirect contact of infected horses with susceptible ones. The observed results demonstrate the circulation of the EIAV in four states in the Northeast region of Brazil.(AU)

Serological and molecular detection of Theileria equi in horses from Sinop, Mato Grosso state, Brazil / Detecção sorológica e molecular de Theileria equi em equinos oriundos de Sinop, Mato Grosso, Brasil

Equine piroplasmosis is the most important tick-borne disease to affect horses in Brazil. Theileria equi is one of the causative agents of equine piroplasmosis. Chronic cases are expected, in which the animals show no apparent signs of infection and remain asymptomatic but constitute a source of the infectious agent that ticks can spread. This study was conducted across 81 ranches located in the municipality of Sinop, State of Mato Grosso, Brazil. A sample calculation was performed to estimate the apparent prevalence of T. equi among horses. A total of 1,853 animals were included in the sampling analysis based on the information available from the Institute of Agricultural and Livestock Defense of Mato Grosso State. The serological analysis of 367 serum samples using an indirect enzyme-linked immunosorbent assay (ELISA) to detect anti-T. equi antibodies revealed that 337 animals were positive, representing a frequency of 90.70%. The molecular analysis to amplify the EMA-1 gene showed positivity in 20 of 89 tested samples. The fragments of four samples were sequenced and analyzed to determine their similarities to sequences from other species, based on sequences deposited at GenBank. All showed 100% similarity with T. equi. Our study represents the first report of T. equi antibodies among the equids in north-central region of Mato Grosso, revealing the widespread distribution of seropositive animals.

A piroplasmose equina é a doença transmitida por carrapatos mais importante em cavalos no Brasil. Theileria equi é um dos agentes causadores da piroplasmose equina. São esperados casos crônicos, nos quais os animais não apresentam sinais aparentes de infecção e permanecem assintomáticos, mas constituem uma fonte de infecção e disseminação por carrapatos. Este estudo foi realizado em 81 fazendas localizadas no município de Sinop, Estado de Mato Grosso, Brasil. Um cálculo amostral foi realizado para estimar a prevalência aparente de T. equi entre cavalos. No total, 1.853 animais foram incluídos na análise amostral com base nas informações disponíveis no Instituto de Defesa Agropecuária do Estado de Mato Grosso. A análise sorológica de 367 amostras de soro por meio de ensaio imunoenzimático indireto (ELISA) para detecção de anticorpos anti-T. equi revelou que 337 animais eram positivos, representando uma frequência de 90,70%. A análise molecular para o gene EMA-1 mostrou positividade em 20 das 89 amostras testadas. Os fragmentos de quatro amostras foram sequenciados e analisados para determinar suas semelhanças com sequências de outras espécies, a partir das sequências depositadas no GenBank. Todos mostraram 100% de similaridade com T. equi. Nosso estudo representa o primeiro relato de anticorpos contra T. equi entre os equídeos na região centro norte de Mato Grosso, revelando a ampla distribuição de animais soropositivos.

Inquérito sorológico de brucelose em machos e fêmeas bovinas em fazendas do Estado de Mato Grosso do Sul / Serological survey of brucellosis in males and females of cattle in the State of Mato Grosso do Sul

Objetivou-se avaliar a ocorrência de animais soro reagentes à brucelose bovina em fazendas localizadas no Estado de Mato Grosso do Sul, por meio de exame sorológico utilizando o Antígeno Acidificado Tamponado (AAT) e discutir as possíveis diferenças entre as soroprevalências de fêmeas e machos. Foram avaliados, a partir do teste de triagem com Antígeno Acidificado Tamponado (AAT), 724 bovinos da raça Nelore, sendo 274 machos e 450 fêmeas, provenientes de oito propriedades com histórico de problemas reprodutivos. O teste foi procedido conforme o protocolo determinado pelo Ministério da Agricultura, Pecuária e Abastecimento (MAPA). Os resultados demonstraram baixa soroprevalência da doença nos bovinos testados, sendo detectada prevalência para a doença de 1,10% nos machos e 2,88% nas fêmeas.Quando se considera o touro isoladamente nos rebanhos, pode-se perceber que a fertilidade é muito mais importante nos machos do que nas fêmeas individualmente, uma vez que os touros podem se acasalar com um número muito maior de fêmeas, seja na monta natural ou na inseminação artificial, demonstrando a importância do inquérito epidemiológico na população geral, principalmente nos machos. A maior frequência da doença foi encontrada nas fêmeas podendo estar relacionada à infecção por Brucella spp. no ambiente decorrente de parto ou aborto tornando as fêmeas transmissoras permanentes da doença.

The objective of this study was to evaluate the occurrence of seroreactive animals to bovine brucellosis in farms located in the State of Mato Grosso do Sul, by means of a serological examination using the Acidified Buffered Antigen (AAT) and to discuss the possible differences between the seroprevalence of females and males. A total of 724 Nellore cattle, 274 males and 450 females, from eight farms with a history of reproductive problems, were evaluated using the screening test with Acidified Buffered Antigen (AAT). The test was carried out according to the protocol determined by the Ministry of Agriculture, Livestock and Food Supply (MAPA).The results showed a low seroprevalence of the disease in the tested cattle, with a prevalence of 1.10% in males and 2.88% in females.When considered the bull alone in herds, it can be shown that fertility is much more important in males than in females individually, since bulls can mate with a much larger number of females, either in natural mating or in artificial insemination, demonstrating the importance of epidemiological survey in the general population, especially in males. The highest frequency of the disease was found in females and may be related to infection by Brucella spp. in the environment from childbirth or abortion making females permanent transmitters of the disease.

Leishmaniose visceral em cães / Visceral leishmaniasis in dogs

Background: Canine visceral leishmaniasis (CVL) is a zoonosis of variable clinical presentation, either in systemic orcutaneous form. Clinical signs include anorexia, ophthalmopathies, and chronic kidney disease. In the state of Santa Catarina, the foci are concentrated in the capital and its adjacencies, in the east side of the state. The objective of this studyis to outline the first three reported cases of CVL in the municipality of Curitibanos, since there are no reports to date inthe region of the mountainous plateau, in the middle west of Santa Catarina.Cases: All dogs were treated at the Veterinary Clinic School of the Federal University of Santa Catarina, Campus Curitibanos. The animals, two males and one female, belonged to the same tutor, resided in Curitibanos, and were attendedbetween 2016 and 2020. In the first case, attended in 2016, the complaint was of eye and skin changes about three monthsago. The animal lived in an urban environment and came from Uruguaiana, Rio Grande do Sul. On physical examination,the animal presented skin peeling, wet and crusty lesions, bloody discharge in the ears and nasal hyperkeratosis, as wellas signs suggestive of uveitis. In this case, euthanasia was carried out. The second case had complaints of respiratory, eyechanges, hyporexia, and polydipsia. The dog was prostrate, dehydrated, with lymphadenomegaly and respiratory disorders, compatible with bacterial pneumonia. Bilateral corneal edema confirmed uveitis. Upon return, the animal remaineddehydrated with enlarged lymph nodes. In the aspiration of the lymph nodes, suggestive forms of Leishmania sp. wereobserved. The recommended confirmatory tests were performed, leading to a definitive diagnosis of CVL. The patientwas treated with miltefosine, but later died. The third case was attended for general evaluation after a positive diagnosisfor CVL during an epidemiological survey of the second case. The animal was alert, tachycardic, and tachypneic with...