Seroprevalence and associated risk factors of Mycoplasma agalactiae and investigation of coinfection with the caprine lentivirus in Rio Grande do Norte, Brazil.

Contagious agalactia is a disease caused by Mycoplasma agalactiae that leads to a reduction or complete stop of milk production. Caprine arthritis encephalitis (CAE) is an infectious disease caused by a lentivirus of the Retroviridae family, member of the small ruminant lentivirus (SRLV) group. Although these diseases are caused by distinct pathogens, the clinical presentation is similar. Hence, this study aimed to perform a serological investigation, as well as to assess correlation between both diseases and risk factors associated in two mesoregions of Rio Grande do Norte, Brazil. Enzyme-linked immunosorbent assay (ELISA) was used for contagious agalactia and western blot for CAE. A total of 538 serum samples were used in this study that were collected from goats and sorted from a blood bank of the Brazilian Agricultural Research Corporation. Seroprevalence of M. agalactiae in flocks from Rio Grande do Norte was 7.8% (42/538). In both regions that were investigated, 25.9% (14/54) of farms had positive animals. CAE results revealed that 3.9% (21/538) of animals and 42.6% (23/54) of farms had this disease. Concerning risk factors, only sex and animal category presented significant relevance (P < 0.05) for contagious agalactia, in which females presented higher frequency of seropositive individuals (10.1%; 39/387). In the animal category, 4.3% (14/326) and 11.1% (36/323) of female breeders were positive for CAE and contagious agalactia, respectively, and significance was identified only in the latter (P < 0.05). In conclusion, there was no correlation between the investigated diseases, considering that no animal demonstrated antibodies for both pathogens.

Seroepidemiological study of Caseous lymphadenitis in sheep from the Northeast region of Brazil using an indirect ELISA.

The objective of this study was to determine the frequency of seropositivity Corynebacterium pseudotuberculosis in sheep in five states of northeastern Brazil, using an indirect enzyme-linked immunosorbent assay (i-ELISA). Young and adult sheep of both sexes were used. Blood samples were collected from 2638 sheep from 223 herds across all states. For the i-ELISA, antigens produced from the strain of C. pseudotuberculosis BRM 029971, a bacterial isolate from the Northeast region of Brazil, were used. Sensitivity and specificity indexes were calculated for the validation of the test, using as reference 49 and 134 serum samples from sheep known to be positive and negative, respectively. The i-ELISA presented four false-negative and four false-positive results, showing a specificity of 97.01%, a sensitivity of 91.84%, and an accuracy of 95.63%. These results were calculated based on an optical density (OD) cutoff point = 0.138. Of the 2638 sheep tested, 996 (37.76%, 95% CI = 35.93-39.62%) were seropositive, and of the 223 evaluated herds, 210 (94.17%, 95% CI = 90.28-96.56%) seropositive. The i-ELISA showed adequate sensitivity and specificity, proving to be a useful tool in the diagnosis of caseous lymphadenitis (CLA) in sheep. Infection by C. pseudotuberculosis, determined by serology, is disseminated in the sheep herds in the states of Northeast Brazil. Thus, there is a need to implement effective control measures that prevent the spread of infectious agents.

Seroepidemiological characterization and risk factors associated with seroconversion to Corynebacterium pseudotuberculosis in goats from Northeastern Brazil.

Goat breeding in the Northeast region of Brazil plays an important socioeconomic role. However, there are significant losses caused by sanitary deficits and infectious diseases, particularly caseous lymphadenitis (CL). Although CL is considered endemic in Northeastern Brazil, a comprehensive and up-to-date study of this disease in goat herds in this region is necessary. The objective of this study was to determine the farm-level and animal-level seroprevalences for the disease and to identify the possible risk factors that characterize CL in the caprine species of five Northeastern's states (Ceará, Piauí, Rio Grande do Norte, Paraíba, and Sergipe). A total of 2744 goat serum samples from 230 farms were collected between 2010 and 2012. The diagnosis of Corynebacterium pseudotuberculosis infection was performed using the indirect ELISA technique. Farm-level and animal-level seroprevalences were 87.8% and 30.3%, respectively, suggesting that C. pseudotuberculosis is widespread in goat herds of the Northeast region. The risk factors were as follows: absence of forage silage (odds ratio = 5.39), not separating animals by sex (odds ratio = 4.16) or by age (odds ratio = 6.30), not replacing old goat breeders (odds ratio = 7.80), and non-treatment of CL lumps prior to spontaneous rupture (odds ratio = 10.34). This study supports the idea that caseous lymphadenitis is widely disseminated in goats from Northeastern Brazil and based on the risk factor analysis attention should be given to the need to establish adequate control measures, such as incision and early drainage of superficial abscesses, quarantine and elimination of affected animals, periodic inspection of the herd, non-introduction of infected animals, and early disposal of animals with recurrent CL.

A panel of protein candidates for comprehensive study of Caprine Arthritis Encephalitis (CAE) infection.

The caprine arthrite encephalite (CAE) is a disease that affects especially dairy goat. The virus shows compartmentalization features, that allows it to hide at certain times during the course of the disease, making it difficult to control. The present study was conducted to identify the major seminal plasma protein profile of goats infected by CAE and its associations with seroconversion using Western blotting. Two groups containing five males each, were used in this experiment. The first group was composed by seropositive animals and the control by seronegative confirmed by Western blotting and PCR. The semen was collected through artificial vagina and after that, two-dimensional electrophoresis and MALDI-TOF MS were used. Seventy-five spots were identified in the goat seminal plasma gels, equivalent to 13 different proteins with more expression. The similar proteins found in both groups and related to reproduction were spermadhesin Z13-like, bodhesin and bodhesin-2, Lipocalin, protein PDC-109-like, and albumin. In infected goats, proteases such as arisulfatase A have been identified, whose function probably is related to metabolism control of sulfatides, involved to virus control. The other ones were bifunctional ATP-dependent dihydroxyacetone kinase/FAD-AMP lyase, cathepsin F isoform X1, disintegrin and metalloproteinase domain-containing protein 2-like isoform X1, clusterin, carbonic anhydrase 2, electron transfer flavoprotein subunit beta, and epididymal secretory glutathione peroxidase. The results of this study show the reaction of the innate immune system against chronic infection of goats by CAE.

In vivo evaluation of the antiretroviral activity of Melia azedarach against small ruminant lentiviruses in goat colostrum and milk.

This study aimed to evaluate in vivo the use of the extract from the leaves of Melia azedarach in the ethyl acetate fraction at a concentration of 150 µg/mL as an antiretroviral treatment against small ruminant lentiviruses (SRLV) in goat colostrum, and milk with a 90-min action. Two groups of six kids were treated with the extract. One group received three supplies of colostrum from does naturally positive for SRLV, treated with the ethyl acetate fraction of M. azedarach (EAF-MA) for three days, while the other group consumed milk from does also carrying the virus with the respective extract twice a day for five days. After undergoing treatment, all animals began to receive thermized milk until weaning (60 days) and were monitored for six months using nested polymerase chain reaction (nPCR) and western blot (WB) tests. The study revealed cumulative percentages of positive animals in WB or nPCR in the milk group of 66.66% on the seventh day, 83.33% in the following week, and 100% at 120 days, while the colostrum group showed values of 66.66% at 14 days, 83.33% at 90 days, and 100% at 120 days. Variation and intermittency were observed in viral detection, but all animals tested positive in WB or nPCR at some point. A potential delay in infection was observed, which was more significant in the colostrum group. The need for the combination of serological and molecular tests for a more efficient detection of the disease is also emphasized.

Risk factors associated with Leptospira sp. seroprevalence in goats in Northeast of Brazil.

Goat farming is of great importance for agriculture in Northeast of Brazil, which owns more than 90 % of the national herd, however, most studies on leptospirosis in goats in this region are local surveys with unplanned sampling. Thus, the objective of this study was to determine the factors associated with the seroprevalence of leptospirosis in goats in Northeast region of Brazil. Blood samples were collected from 4121 goats from 222 herds, belonging to the states of Ceará, Paraíba, Piauí , Rio Grande do Norte and Sergipe. Serological diagnosis was carried out using the microscopic agglutination test (MAT). In 82.9 % (95 % CI = 77.3-87.2 %) of the herds there was at least one animal seropositive for at least one of the serogroups of Leptospira sp., and 18.73 % (95 % CI = 17.5-19.9 %) of the animals were seropositive. Antibody titers ranged from 100 to 800 and the most frequent serogroups were Autumnalis (23.06 %) and Australis (20.98 %). According to the robust Poisson regression analysis, four factors were associated with seroprevalence: animal sex (female) (P = 0.040), educational level of the farmer (elementary school) (P < 0.001), contacting institution for technical support (P = 0.040) and presence of irrigation area (P = 0.002). There was a high frequency of Leptospira sp. in goats in the states of Northeast region of Brazil, and it's suggested the need for better technological and preventive assistance of herds.

Detection and isolation of small ruminant lentivirus in the amniotic fluid of goats.

The objective of this study was to verify the presence of small ruminant lentivirus in the amniotic fluid of goats using molecular tests and viral isolation by cocultivation in the amniotic fluid of naturally infected goats. The study analyzed eight goats: seven were small ruminant lentivirus-positive and one was negative. The amniotic fluid was collected from each of the eight animals during cesarean section at 147 days of pregnancy. Cocultivation was undertaken using secondary goat nictitating membrane cell cultures obtained by explant from a small ruminant lentivirus-negative calf followed by trypsinization and sub-cultivation of the cells for 63 days. During this period, five supernatant collections were performed for DNA extraction and subsequent nested polymerase chain reaction. DNA was extracted from the amniotic fluid after 3 h of cellular sedimentation, from which a sample of 600 µL was taken from the sediment and another 600 µL sample from the supernatant. After DNA extraction, nested polymerase chain reaction was performed. Of the eight goats, 62.5 % (05/08) were small ruminant lentivirus-positive, with 43.75 % (07/16) of the total samples positive when considering the two repetitions (supernatant and cell sediment). Moreover, positivity was confirmed by small ruminant lentivirus pro-viral DNA amplification in the cell supernatant throughout the cocultivation period. Small ruminant lentivirus were present in the amniotic fluid samples from the naturally infected goats indicating an intrauterine transmission route. Moreover, this biological fluid can be adopted for the diagnosis of these lentiviruse because it is an important risk factor related to intrauterine transmission.

Factors associated with the seroprevalence of caseous lymphadenitis in sheep from Northeastern Brazil.

Caseous lymphadenitis (CL) is one of the main infectious diseases in sheep, responsible for great economic losses, mainly in Northeast region of Brazil, which has the largest sheep flock in the country. Thus, the aim of this survey was to determine the factors associated with the seroprevalence of this disease in sheep from Northeastern Brazil using a planned sample of flocks and animals. Samples were collected from 2312 adult and young sheep of both sex from 196 farms, located in 51 municipalities in five Northeastern Brazilian states (Ceará, Paraíba, Piauí, Rio Grande do Norte, and Sergipe). Serological diagnosis was performed using the indirect ELISA technique. Factors associated with seropositivity were identified by variables extracted from epidemiological questionnaires administered to breeders, using univariable and multivariable analyses. Factors associated with prevalence were purebred sheep (Prevalence ratio - PR = 1.189; P = 0.017), ram acquisition from animal expositions (PR = 1.192; P = 0.020), offspring is separated from ewes (PR = 1.132; P = 0.048), water supplied to sheep from ponds (PR = 1.365; P = 0.002), and delayed disposal of infected animals (PR = 1.263; P = 0.027). Flock-level and animal-level prevalences were 93.88 % (95 % CI = 89.60-96.46%) and 37.46 % (95 % CI = 35.51-39.45%), respectively. In view of the high prevalence found here and based on the associated factors, the implementation of an efficient CL control programme in Northeastern Brazil is recommended.

Vertical transmissibility of small ruminant lentivirus.

This study aimed to evaluate by means of Nested Polymerase Chain Reaction (nPCR), co-cultivation and sequencing, with genetic comparison between strains (mother/newborn), the occurrence of vertical transmission of Small Ruminant Lentiviruses (SRLV) from naturally occurring nannies infected for their offspring. For the detection of SRLV seropositive progenitors, blood was collected from 42 nannies in the final third of gestation in tubes with and without anticoagulant. The diagnostic tests used were Western Blot (WB) and nPCR. During the period of birth, the same blood collection procedure was performed on 73 newborns at zero hours of birth, with the same diagnostic tests. Seventeen blood samples from seven-day-old kids, proven positive for SRLV by nPCR, chosen at random, were subjected to coculture in goat synovial membrane (GSM) cells for 105 days. The pro-viral DNA extracted from the cell supernatant from the coculture was subjected to nPCR. For DNA sequencing from the nPCR products, nine positive samples were chosen at random, four nannies with their respective offspring, also positive. Each sample was performed in triplicate, thus generating 27 nPCR products of which only 19 were suitable for analysis. Among the 42 pregnant goats, in 50% (21/42) pro-viral DNA was detected by nPCR, while in the WB, only 7.14% (3/42) presented antibodies against SRLV. Regarding neonates, of the 73 kids, 34 (46.57%) were positive for the virus, using the nPCR technique, while in the serological test (WB), three positive animals (4.10%) were observed. The coculture of the 17 samples with a positive result in the nPCR was confirmed in viral isolation by amplification of the SRLV pro-viral DNA. When aligned, the pro-viral DNA sequences (nannies and their respective offspring) presented homology in relation to the standard strain CAEV Co. It was concluded that the transmission of SRLV through intrauterine route was potentially the source of infection in the newborn goats.

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.

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...

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)

Risk factors associated with seroprevalence of Chlamydia abortus in sheep farms in Ceará, Brazil

Background: Chlamydia abortus infections (Chlamydiosis) can cause reproductive problems in sheep, such as abortionsand birth defects, leading to farm productivity loss. The symptoms, which are similar to other reproductive diseases, andthe microbial pathogenesis make the clinical diagnosis difficult. Chlamydia abortus is a zoonotic pathogen, making it apublic health issue because it can infect and induce abortions in humans. This study investigated anti-C. abortus antibodylevels and infection risk factors in sheep in the State of Ceará, Brazil.Materials, Methods & Results: Forty-three properties from 10 municipalities in 4 mesoregions in the State of Ceará, Brazil(Sertões, metropolitan Fortaleza, North Ceará and Northwest Ceará) with sheep, goats, cattle, and horses were visited.Five hundred and four serological samples from sheep were collected and tested for anti-C. abortus antibodies using anEnzyme-linked Immunosorbent Assay (ELISA) [IDEXX®, Australia] and all procedures were performed in accordancewith the manufacturer’s instructions at the Clinical Pathology Laboratory of EMBRAPA Goats and Sheep (Sobral, Brazil). Individual questionnaires were completed about sheep breeding practices and to identify possible C. abortus risks.Seropositive results were found in 18.45 % (93/504 individuals) of sheep, and 88.37 % (38/43 properties) of the herds hadat least one seropositive animal. The number of seropositive individuals was significantly different between adults andewes [P < 0.01; Odds Ratio (OR) = 0.510; 95% confidence interval (CI) = 0.306 - 0.850]. Logistic regression modelingidentified a missing health certificate request for newly acquired animals as a chlamydiosis risk factor [P = 0.038; OR =2.672; 95% CI = 1.058 - 6.749].Discussion: The prevalence of anti-C. abortus...

Spatial distribution of serologically reactive sheep to Leptospira spp. in the northeast region of Brazil

Considering the importance of leptospirosis in sheep farming and public health and the significance of identifying which serogroups circulate in sheep within each region, the objective of this study was to determine the seroprevalence and spatial distribution of the most frequent serogroups causing infection by Leptospira sp. in ovine herds in the Northeast region of Brazil. Blood samples were collected from 4197 sheep from 229 herds in 7 Northeastern States. Sera were analyzed via microscopic agglutination test (MAT). The frequency of seroreactive sheep for Leptospira sp. was 14.06%. The states of Alagoas, Ceará, Paraíba, Piauí, Rio Grande do Norte, and Sergipe, located in the Caatinga biome, had the highest frequencies of serologically reactive sheep, and Maranhão, in the Cerrado biome, had the lowest frequency. The most frequent serogroups were Autumnalis (19.49%), Australis (15.76%) and Serjoe (14.41%). In the states of Ceará, Paraíba, Rio Grande do Norte, and Sergipe, 100% of their municipalities had at least one seroreactive animal. The highest frequencies of seropositive animals were found in the municipalities of União (50%), Passagem (49.06%), Canindé (48.89%), Igaci (28.95%), Gararu (31.2%), Pirapemas (17.5%), and Angicos (16%) located in the states of Piauí, Paraíba, Ceará, Alagoas, Sergipe, Maranhão and Rio Grande do Norte, respectively. The animal-level prevalence (14.06%) obtained in the present study is significant, especially considering the rustic nature of the species and the adverse conditions of the region for the infectious agent. In semi-arid conditions, it has been suggested that perhaps sheep do not seroconvert detectable titers on MAT with a cut-off point of 1:100. It is important to highlight that the ovine population in the Northeast region of Brazil is composed of mixed animals, which have been considered more resistant to infection by Leptospira spp. Also, environmental factors hostile to the survival of the infectious agent in the studied region should be taken into consideration, since they may have influenced the seropositive animal-level prevalence. A noteworthy variation was observed in agglutinin titers, which ranged from 100 to 1,600, where 80.2% of the positive samples had titers ≤ 200. It is important to highlight that more elevated titers (≥ 400) were obtained in all seven states, which may suggest an acute infection caused by a non-adapted serovar, indicating that preventive and control measures focused on possible infection sources for sheep should be adopted. Although some states showed the same serogroups as the most frequent, a variety of serogroups was observed in municipalities, which may indicate different sources of infection, whether interspecies, intraspecies, or via alternative routes of transmission in semi-arid conditions, such as venereal. This indicates that even though sheep are more resistant to infection, they become exposed due to the environment or management conditions. As such, identification, isolation, and treatment of the affected animals are alternative measures recommended for prevention and control of leptospirosis in sheep in the semi-arid region. It is evident that despite the lack of rain observed in the last decade in the Northeast region of Brazil, which prevented the formation of favorable environments for the presence of Leptospira, the infectious agent remains among the sheep, as well as other production and wild animals in the region. Some factors may be contributing to this scenario, such as the fact that sheep farming in the region is = characterized mainly by subsistence systems, where veterinary assistance and adequate sanitary management are absent, thus increasing the possibility of contact with Leptospira. (AU)

Risk factors associated with seroprevalence of Chlamydia abortus in sheep farms in Ceará, Brazil

Background: Chlamydia abortus infections (Chlamydiosis) can cause reproductive problems in sheep, such as abortionsand birth defects, leading to farm productivity loss. The symptoms, which are similar to other reproductive diseases, andthe microbial pathogenesis make the clinical diagnosis difficult. Chlamydia abortus is a zoonotic pathogen, making it apublic health issue because it can infect and induce abortions in humans. This study investigated anti-C. abortus antibodylevels and infection risk factors in sheep in the State of Ceará, Brazil.Materials, Methods & Results: Forty-three properties from 10 municipalities in 4 mesoregions in the State of Ceará, Brazil(Sertões, metropolitan Fortaleza, North Ceará and Northwest Ceará) with sheep, goats, cattle, and horses were visited.Five hundred and four serological samples from sheep were collected and tested for anti-C. abortus antibodies using anEnzyme-linked Immunosorbent Assay (ELISA) [IDEXX®, Australia] and all procedures were performed in accordancewith the manufacturers instructions at the Clinical Pathology Laboratory of EMBRAPA Goats and Sheep (Sobral, Brazil). Individual questionnaires were completed about sheep breeding practices and to identify possible C. abortus risks.Seropositive results were found in 18.45 % (93/504 individuals) of sheep, and 88.37 % (38/43 properties) of the herds hadat least one seropositive animal. The number of seropositive individuals was significantly different between adults andewes [P < 0.01; Odds Ratio (OR) = 0.510; 95% confidence interval (CI) = 0.306 - 0.850]. Logistic regression modelingidentified a missing health certificate request for newly acquired animals as a chlamydiosis risk factor [P = 0.038; OR =2.672; 95% CI = 1.058 - 6.749].Discussion: The prevalence of anti-C. abortus...(AU)

Efficacy of measures to control caprine arthritis-encephalitis in dairy herd with high clinical and serological prevalence / Eficácia das medidas de controle da artrite-encefalite caprina em rebanho leiteiro com alta prevalência clínica e sorológica

This study examined the effectiveness of control measures for caprine arthritis-encephalitis in a herd with 431 dairy goats in an intensive rearing system. All animals older than six months were initially tested by agar gel immunodiffusion (AGID) and separated into seropositive and seronegative. Control measures were implemented for two years and ten months. Five serological examinations were subsequently performed—two by AGID and three by the Western Blot (WB) technique. In these tests, animals that tested negative in the previous serological examination were evaluated along with those older than six months which had not yet been examined. The effectiveness of control was evaluated based on the incidence of the disease. Seroconverted animals were stratified according to age, physiological status and dam serology. For the effect of time, logistic regression was performed at the 5% significance level, with values converted into likelihood. General incidence and incidence as a function of age and physiological status were evaluated by analysis of variance, with means compared by Tukeys test at 5% significance. The ratio test was used for incidence and physiological status, and the agreement between the AGID and WB tests was determined by the Kappa coefficient. Animals that seroconverted and were born to positive dams were compared with those born to dams negative at birth by the Chi-square test, and the same was applied for the number of discarded animals. Initially, 54.24% (179/330) positive and 257 seroconverted animals were identified after the start of control. Higher incidence occurred in the animal saged between 13 and 36 months and in lactating does. Seroconversions among offspring of seropositive dams were higher than in the offspring of seronegative dams (p < 0.001). High infection rates were identified in the sires. The obtained [...].(AU)

Avaliou-se a eficácia das medidas de controle para a artrite-encefalite caprina em rebanho com 431 caprinos leiteiros em regime intensivo de criação. Todos os animais com idade superior a seis meses foram inicialmente testados por imunodifusão em gel de ágar (IDGA) e soropositivos e soronegativos separados. Durante dois anos e dez meses medidas de controle foram instituídas. Cinco testes sorológicos foram posteriormente realizados, dois por IDGA e três por Western Blot (WB). Nesses testes eram avaliados os animais negativos na sorologia anterior e acrescidos os com mais de seis meses, ainda não avaliados. A eficácia do controle foi avaliada pela incidência da enfermidade. Animais que soroconverteram foram estratificados quanto a idade, estado fisiológico e sorologia das progenitoras. Para o efeito do tempo foi realizado a regressão logística a 5% de significância, convertidos em razão de probabilidades. A incidência geral e incidência em função da idade e estado fisiológico foram avaliadas pela análise variância, comparando as médias pelo teste Tukey a 5 % de significância. O teste de proporções foi utilizado para incidência e estado fisiológico, e a concordância entre os testes IDGA e Wb realizada através do coeficiente Kappa. Os animais que soroconverteram e eram nascidos de progenitoras positivas foram comparados com aqueles de progenitoras negativas ao parto pelo teste de Qui-quadrado, assim como o número de animais descartados. Inicialmente identificou-se 54,24%(179/330) de animais positivos e 257 soroconverteram após início do controle. Incidências maiores ocorreram nos animais entre 13 e 36 meses e nas lactantes. Soroconversões em crias de progenitoras soropositivas foram maiores que nas de progenitoras soronegativas (p < 0,001). Altas taxas de infecção foram identificadas nos reprodutores. Os resultados obtidos não foram satisfatórios, pois as medidas não contribuíram para evitar novos casos, demonstrando que existem momentos de infecção que [...].(AU)

Efficacy of measures to control caprine arthritis-encephalitis in dairy herd with high clinical and serological prevalence / Eficácia das medidas de controle da artrite-encefalite caprina em rebanho leiteiro com alta prevalência clínica e sorológica

This study examined the effectiveness of control measures for caprine arthritis-encephalitis in a herd with 431 dairy goats in an intensive rearing system. All animals older than six months were initially tested by agar gel immunodiffusion (AGID) and separated into seropositive and seronegative. Control measures were implemented for two years and ten months. Five serological examinations were subsequently performed—two by AGID and three by the Western Blot (WB) technique. In these tests, animals that tested negative in the previous serological examination were evaluated along with those older than six months which had not yet been examined. The effectiveness of control was evaluated based on the incidence of the disease. Seroconverted animals were stratified according to age, physiological status and dam serology. For the effect of time, logistic regression was performed at the 5% significance level, with values converted into likelihood. General incidence and incidence as a function of age and physiological status were evaluated by analysis of variance, with means compared by Tukey’s test at 5% significance. The ratio test was used for incidence and physiological status, and the agreement between the AGID and WB tests was determined by the Kappa coefficient. Animals that seroconverted and were born to positive dams were compared with those born to dams negative at birth by the Chi-square test, and the same was applied for the number of discarded animals. Initially, 54.24% (179/330) positive and 257 seroconverted animals were identified after the start of control. Higher incidence occurred in the animal saged between 13 and 36 months and in lactating does. Seroconversions among offspring of seropositive dams were higher than in the offspring of seronegative dams (p < 0.001). High infection rates were identified in the sires. The obtained [...].

Avaliou-se a eficácia das medidas de controle para a artrite-encefalite caprina em rebanho com 431 caprinos leiteiros em regime intensivo de criação. Todos os animais com idade superior a seis meses foram inicialmente testados por imunodifusão em gel de ágar (IDGA) e soropositivos e soronegativos separados. Durante dois anos e dez meses medidas de controle foram instituídas. Cinco testes sorológicos foram posteriormente realizados, dois por IDGA e três por Western Blot (WB). Nesses testes eram avaliados os animais negativos na sorologia anterior e acrescidos os com mais de seis meses, ainda não avaliados. A eficácia do controle foi avaliada pela incidência da enfermidade. Animais que soroconverteram foram estratificados quanto a idade, estado fisiológico e sorologia das progenitoras. Para o efeito do tempo foi realizado a regressão logística a 5% de significância, convertidos em razão de probabilidades. A incidência geral e incidência em função da idade e estado fisiológico foram avaliadas pela análise variância, comparando as médias pelo teste Tukey a 5 % de significância. O teste de proporções foi utilizado para incidência e estado fisiológico, e a concordância entre os testes IDGA e Wb realizada através do coeficiente Kappa. Os animais que soroconverteram e eram nascidos de progenitoras positivas foram comparados com aqueles de progenitoras negativas ao parto pelo teste de Qui-quadrado, assim como o número de animais descartados. Inicialmente identificou-se 54,24%(179/330) de animais positivos e 257 soroconverteram após início do controle. Incidências maiores ocorreram nos animais entre 13 e 36 meses e nas lactantes. Soroconversões em crias de progenitoras soropositivas foram maiores que nas de progenitoras soronegativas (p < 0,001). Altas taxas de infecção foram identificadas nos reprodutores. Os resultados obtidos não foram satisfatórios, pois as medidas não contribuíram para evitar novos casos, demonstrando que existem momentos de infecção que [...].

Study of metalloproteinases in the blood of goats experimentally infected with caprine encephalitis arthritis virus / Estudo de metaloproteinases no sangue de caprinos experimentalmente infectados pelo vírus da artrite encefalite caprina

Caprine arthritis encephalitis is a lentiviral disease that leads to considerable losses in goat farming. In the acute phase of viral infection, though antiviral antibodies are produced by the host's immune system, they are not sufficient to be detected by serological tests. Acute infections begin with an incubation period, during which the viral genome replicates and host innate responses are initiated. Matrix metalloproteinases (MMPs) are enzymes that play an important role in the physiological and pathological processes of tissue remodeling. The present study aimed to evaluate the expression of MMPs and their activity in the blood serum of male goats experimentally infected with caprine arthritis encephalitis virus (CAEV). Five dairy male goats, aged 3-4 years, were intravenously inoculated with CAEV Cork strain (titer: 105-6 TCID50/mL) after being tested negative for CAEV thrice at consecutive intervals of 30 days using western blot analysis and nested-PCR. The study included three stages: S1 or pre-infection stage; S2 or seroconversion stage, corresponding to the occurrence of first seroconversion; and S3 or post-seroconversion stage, corresponding to 23 weeks after seroconversion. Zymography was performed for the samples using gelatin zymography gels (12.5%), which were subjected to electrophoresis at 170V, 1A, and 300W for 50-70 min. The density of MMP-2 was found to be lower at S1 (1456.20 pixels) than that at S2 and S3 (1943.80 and 2104.40 pixels, respectively) (P < 0.05); and the density of MMP-9 was found to be lower at S3 (133.60 pixels) than that at S1 and S2 (359.60 and 370.60 pixels, respectively). The density of proMMP-2 was low at S1 and S3 (130.45 and 145.20 pixels, respectively). On the other hand, the density of proMMP-9 was statistically different between S1 and S3 (89.22 vs. 415.60 pixels). Both proMMP-2 and proMMP-9 were absent at S2. Thus, MMP-2 and MMP9 exhibited opposite behaviors depending on the stage [...].

A encefalite por artrite caprina é uma doença lentiviral que leva a perdas consideráveis na criação de caprinos. Na fase aguda da infecção viral, embora os anticorpos antivirais sejam produzidos pelo sistema imunológico do hospedeiro, eles não são suficientes para serem detectados por testes sorológicos. As infecções agudas começam com um período de incubação, durante o qual o genoma viral se replica e as respostas inatas do hospedeiro são iniciadas. As metaloproteinases da matriz (MMPs) são enzimas que desempenham um papel importante nos processos fisiológicos e patológicos da remodelação tecidual. O presente estudo teve como objetivo avaliar a expressão de MMPs e sua atividade no soro sanguíneo de reprodutores caprinos infectados experimentalmente pelo vírus da encefalite por artrite caprina(CAEV). Cinco machos caprinos, com idades entre 3-4 anos, foram inoculadas por via intravenosa com a cepa de CAEV Cork (título: 105-6 TCID50/mL) após serem testados negativamente para CAEV três vezes em intervalos consecutivos de 30 dias usando a análise de western blot e nested-PCR. O estudo incluiu três etapas: estágio S1 ou pré-infecção; S2 ou estágio de soroconversão, correspondente à ocorrência de primeira soroconversão; e S3 ou pós-soroconversão, correspondendo a 23 semanas após a soroconversão. A zimografia foi realizada para as amostras utilizando gel de gelatina (12,5%), que foram submetidos à eletroforese em 170V, 1A e 300W por 50-70 min. Verificou-se que a densidade de MMP-2era menor em S1 (1456,20 pixels) do que em S2 e S3 (1943,80 e 2104,40 pixels, respectivamente) (P <0,05); e a densidade de MMP-9 foi menor em S3 (133,60 pixels) do que em S1 e S2 (359,60 e 370,60 pixels, respectivamente). A densidade do proMMP-2 era baixa em S1 e S3 (130,45 e 145,20 pixels, respectivamente). Por outro lado, a densidade do proMMP-9 foi estatisticamente diferente entre S1 e S3(89,22 vs. 415,60 pixels). ProMMP-2 e proMMP-9 estavam ausentes no S2. Assim, MMP-2 e [...].

Study of metalloproteinases in the blood of goats experimentally infected with caprine encephalitis arthritis virus / Estudo de metaloproteinases no sangue de caprinos experimentalmente infectados pelo vírus da artrite encefalite caprina

Caprine arthritis encephalitis is a lentiviral disease that leads to considerable losses in goat farming. In the acute phase of viral infection, though antiviral antibodies are produced by the host's immune system, they are not sufficient to be detected by serological tests. Acute infections begin with an incubation period, during which the viral genome replicates and host innate responses are initiated. Matrix metalloproteinases (MMPs) are enzymes that play an important role in the physiological and pathological processes of tissue remodeling. The present study aimed to evaluate the expression of MMPs and their activity in the blood serum of male goats experimentally infected with caprine arthritis encephalitis virus (CAEV). Five dairy male goats, aged 3-4 years, were intravenously inoculated with CAEV Cork strain (titer: 105-6 TCID50/mL) after being tested negative for CAEV thrice at consecutive intervals of 30 days using western blot analysis and nested-PCR. The study included three stages: S1 or pre-infection stage; S2 or seroconversion stage, corresponding to the occurrence of first seroconversion; and S3 or post-seroconversion stage, corresponding to 23 weeks after seroconversion. Zymography was performed for the samples using gelatin zymography gels (12.5%), which were subjected to electrophoresis at 170V, 1A, and 300W for 50-70 min. The density of MMP-2 was found to be lower at S1 (1456.20 pixels) than that at S2 and S3 (1943.80 and 2104.40 pixels, respectively) (P < 0.05); and the density of MMP-9 was found to be lower at S3 (133.60 pixels) than that at S1 and S2 (359.60 and 370.60 pixels, respectively). The density of proMMP-2 was low at S1 and S3 (130.45 and 145.20 pixels, respectively). On the other hand, the density of proMMP-9 was statistically different between S1 and S3 (89.22 vs. 415.60 pixels). Both proMMP-2 and proMMP-9 were absent at S2. Thus, MMP-2 and MMP9 exhibited opposite behaviors depending on the stage [...].(AU)

A encefalite por artrite caprina é uma doença lentiviral que leva a perdas consideráveis na criação de caprinos. Na fase aguda da infecção viral, embora os anticorpos antivirais sejam produzidos pelo sistema imunológico do hospedeiro, eles não são suficientes para serem detectados por testes sorológicos. As infecções agudas começam com um período de incubação, durante o qual o genoma viral se replica e as respostas inatas do hospedeiro são iniciadas. As metaloproteinases da matriz (MMPs) são enzimas que desempenham um papel importante nos processos fisiológicos e patológicos da remodelação tecidual. O presente estudo teve como objetivo avaliar a expressão de MMPs e sua atividade no soro sanguíneo de reprodutores caprinos infectados experimentalmente pelo vírus da encefalite por artrite caprina(CAEV). Cinco machos caprinos, com idades entre 3-4 anos, foram inoculadas por via intravenosa com a cepa de CAEV Cork (título: 105-6 TCID50/mL) após serem testados negativamente para CAEV três vezes em intervalos consecutivos de 30 dias usando a análise de western blot e nested-PCR. O estudo incluiu três etapas: estágio S1 ou pré-infecção; S2 ou estágio de soroconversão, correspondente à ocorrência de primeira soroconversão; e S3 ou pós-soroconversão, correspondendo a 23 semanas após a soroconversão. A zimografia foi realizada para as amostras utilizando gel de gelatina (12,5%), que foram submetidos à eletroforese em 170V, 1A e 300W por 50-70 min. Verificou-se que a densidade de MMP-2era menor em S1 (1456,20 pixels) do que em S2 e S3 (1943,80 e 2104,40 pixels, respectivamente) (P <0,05); e a densidade de MMP-9 foi menor em S3 (133,60 pixels) do que em S1 e S2 (359,60 e 370,60 pixels, respectivamente). A densidade do proMMP-2 era baixa em S1 e S3 (130,45 e 145,20 pixels, respectivamente). Por outro lado, a densidade do proMMP-9 foi estatisticamente diferente entre S1 e S3(89,22 vs. 415,60 pixels). ProMMP-2 e proMMP-9 estavam ausentes no S2. Assim, MMP-2 e [...].(AU)

Assessment of caseous lymphadenitis in goats in a slaughterhouse in the Brazilian semi-arid region and estimates of economic losses due to carcass condemnation / Avaliação da linfadenite caseosa em caprinos em um abatedouro na região do semiárido brasileiro e estimativas de perdas econômicas por condenação de carcaças

Caseous lymphadenitis (CLA) caused by the pathogenic bacterium, Corynebacterium pseudotuberculosis, results in economic loss in goat farming. Its prevalence in Brazilian herds varies from 5 to 50%, generating expenses with treatment, loss in production, carcass and organ condemnation, and reduced price of the hide. This study aimed to describe the distribution of lesions and quantify the loss associated with CLA in goat breeding due to condemnation in a slaughterhouse located in a semiarid region. The study was conducted at the municipal slaughterhouse of the city of Patos, State of Paraíba, Northeastern Brazil, where goats from this intermediate geographical region are slaughtered. In 2017, 3,662 animals were slaughtered, an average of 305 per month. During the study period, from March to August 2017, 304 goats of both sexes and different ages of undefined breed were assessed. In the ante-mortem examination, inspection and palpation of the superficial lymph nodes was done; in the post-mortem examination, organ and viscera were assessed. The caseous material collected was sent for microbiological analysis. Of the 304 individuals, 227 [74.67% (95% CI: 69.50-79.23)] did not have any lesions, while 77 [25.33% (95% CI: 20.77-30.50)] showed abscesses suggestive of CLA, which was confirmed in 65 goats [84.41% (95% CI: 76.30-92.50)]. Sex (P = 0.044) and age (P = 0.002) were associated with infection. While the sex of the animals affected carcass and viscera weights and carcass yield, their age affected live weight, carcass, organ, and viscera weights; conversely, the incidence of CLA did not affect these variables. The average price in Real (R$) of a kilogram (kg) of goat meat cuts and “offal” was used to calculate the economic loss. Considering the average carcass weight (11,485 kg), organ weight (1,085 kg), and viscera weight (2,013 kg), 4,433.232 kg of meat was produced, resulting in R$ 72,945.43. The condemnations resulted in the loss of 269.894 kg, which is equivalent to R$ 4,540.33. CLA is responsible for a negative impact of 6.09% on production. Implementing control measures for this disease is important to increase the financial return in goat farming.

Linfadenite Caseosa (LC) causada pela bactéria patogênica Corynebacterium pseudotuberculosis resulta em perda econômica na caprinocultura. No rebanho brasileiro sua prevalência varia de 5 a 50%, gerando gastos com tratamento, queda de produção, condenação de carcaça e órgãos, e desvalorização da pele. O objetivo deste trabalho foi quantificar a perda associada à LC na caprinocultura de corte por condenação em abatedouro do semiárido, bem como caracterizar a distribuição das lesões. O trabalho foi realizado no Abatedouro Municipal da cidade de Patos, Estado da Paraíba, Nordeste do Brasil, onde ocorre abate de caprinos provenientes desta região geográfica intermediária. No ano de 2017 foram abatidos 3.662 animais, uma média de 305 por mês. Durante o período de estudo, março a agosto do mesmo ano, avaliou-se 304 caprinos sem padrão racial definido, ambos os sexos e idades diversas. No exame ante mortem foram submetidos à inspeção e palpação de linfonodos superficiais; post mortem, órgãos e vísceras. O material caseoso coletado foi submetido à análise microbiológica para diagnóstico. Dos 304 indivíduos, 227 [74,67% (IC 95%: 69,50-79,23)] não apresentaram qualquer lesão, enquanto 77 [25,33% (IC 95%: 20,77-30,50)] evidenciaram abscessos sugestivos de LC, com confirmação em 65 [84,41% (IC 95%: 76,30-92,50)]. Constatou-se associação de sexo (P = 0,044) e idade (P = 0,002) à infecção. Foi verificado efeito do sexo dos animais nas variáveis peso de carcaça, peso de vísceras e rendimento de carcaça, bem como houve efeito da idade nas variáveis peso vivo, peso de carcaça, peso de órgãos e peso de vísceras; no entanto, não houve efeito da ocorrência de LC nessas variáveis. Para cálculo de perda econômica adotou-se o preço médio em Real (R$) do quilograma (kg) dos cortes de carne caprina, e “miúdos”. Considerando o peso médio de carcaça (11,485 kg), órgãos (1,085 kg) e vísceras (2,013 kg), houve produção de 4.433,232 kg, resultando em R$ 72.945,43. As condenações subtraíram 269,894 kg, equivalente a R$ 4.540,33. Atribui-se à LC um impacto negativo de 6,09% na produção. Implementar medidas de controle para essa doença é importante para que se amplie o retorno financeiro à atividade.