A. Andrioli

Fatores de risco na transmissão do lentivírus caprino pelo sêmen

The objective of this work was to evaluate the presence of the DNA provirus of the caprine lentivirus (LVC) in ejaculates of naturally infected males, and to verify the influence of the wash of the semen as well as the presence of testicle inflammation on the viral load. Eight semen collections of seven soropositive reproducers were accomplished, four before testicle injury and four after injury. Amongst the collections carried out at the same week, in one the ejaculate was washed, to withdraw the plasma seminal, and in the other it was not. The provirus DNA was identified both by Nested polymerase chain reaction technique (Nested PCR) and by the viral isolation. The virus was isolated in 7.1% of the samples. The PCR identified the provirus DNA in 35.7% of all samples, 17.9% in the washed samples and 53.6% of the integral semen samples. The injury of the testicle tends to greater flow of virus for the semen, therefore, before injury, 21.4% of the samples were positive and after-injury, 50%. Risk of transmission of the LVC by semen of goat reproducers is strengthened by the presence of testicle inflammations and the fact that the criopreserved semen contains the LVC in infecting form.

Interspecific transmission of small ruminant lentiviruses from goats to sheep

This study was conducted in order to evaluate the transmission of caprine lentivirus to sheep using different experimental groups. The first one (colostrum group) was formed by nine lambs receiving colostrum from goats positive for small ruminant lentiviruses (SRLV). The second group (milk group) was established by nine lambs that received milk of these goats. Third was a control group, consisting of lambs that suckled colostrum and milk of negative mothers. Another experimental group (contact group) was formed by eight adult sheep, confined with two naturally infected goats. The groups were monitored by immunoblotting (IB), enzyme-linked immunosorbent assay (ELISA), agar gel immunodiffusion (AGID) and nested polymerase chain reaction (nPCR). All lambs that suckled colostrum and milk of infected goats and six sheep of the contact group had positive results in the nPCR, although seroconversion was detected only in three of the exposed animals, with no clinical lentiviruses manifestation, in 720 days of observation. There was a close relationship between viral sequences obtained from infected animals and the prototype CAEV-Cork. Thus, it was concluded that SRLV can be transmitted from goats to sheep, however, the degree of adaptation of the virus strain to the host species probably interferes with the infection persistence and seroconversion rate.

Padronização do Elisa indireto e Western Blot para diagnóstico da artrite-encefalite caprina

Caprine arthritis-encephalitis (CAE) is routinely diagnosed with the Agarose Gel Immunodiffusion (AGID) technique, which is considered to have low sensitivity. The objective of this study was to standardize testing i-Elisa and Western Blot for early detection of antibodies against CAEV in goats and compare the results obtained in these tests with proof of AGID. For standardization of i-Elisa and WB, different concentrations and dilutions of antigen, sera and conjugate were used. In the i-Elisa, rigid microplate with 96 wells was adopted, and the combination that showed the best result was a concentration of 0.5µg/ well of antigen and dilutions of the serum of 1:100 and conjugate of 1:1500. In the WB nitrocellulose membranes were used, and the dilutions of the serum were defined at 1:50 and conjugate at 1:15000. To evaluate the performance of the techniques, 222 goat serum samples were tested and the data were compared with the AGID. The sensitivity and specificity of Elisa-i/IDGA, WB/AGID and WB/Elisa-i were 70% and 91%, 100% and 72.6%, 84.6% and 76.5%, concomitantly. The Kappa index of these tests was 0.35, 0.2 and 0.36, respectively. The i-Elisa and WB techniques were more sensitive than the AGID and can be used as tools for early diagnosis of CAE.

Detecção do vírus da Artrite Encefalite Caprina por nested PCR e nested RT-PCR em ovócitos e fluido uterino

Caprine arthritis-encephalitis (CAE) is an infectious disease caused by the caprine arthritis-encephalitis virus (CAEV), belonging to the lentivirus genus. The presence of the virus has been observed in the nervous system, respiratory tract and mammary gland, and also in the male and female genital tract. The objective of this study is to identify the virus in oocyte and uterine fluid of infected goats by molecular diagnostic techniques, in order to assess the possibility of CAEV transmission with reproduction. Thirteen infected goats were selected and submitted to euthanasia for the collection of the reproductive system, aspiration of the uterine fluid and dissection of ovaries for oocyte collection. In order to identify the CAEV in the collected material, in the protovirus and free forms, it was submitted to the nRT-PCR and nPCR techniques, respectively. As a result, it was observed that 53.8% of oocytes were positive to nRT-PCR, while only 9.1% were positive to nPCR. The nRT-PCR also identified the virus in the uterine fluid of 46.1% of the tested females. Even though the 13 goats had CAEV, 30.8% presented negative results in nPCR and nRT-PCR in all of the analyzed samples (oocyte and uterine fluid). This work concludes that nRT-PCR and nPCR can be used in the diagnosis of CAE for the analysis with oocytes and uterine fluid, and that the presence of CAEV in these materials points out to the risk of CAEV transmission through reproductive technologies used in females.

Eficiência da recuperação de embriões e os efeitos de consecutivas colheitas sobre o aparelho reprodutor de doadoras da espécie caprina

The purpose of this trial was to compare the efficiency and effect of consecutive embryo recoveries by three different methods (T1 - transcervical; T2 laparoscopy and T3 laparotomy) on the reproductive activity of goat donors. Ten goats were allocated into each treatment (T1, T2 and T3) and submitted to three consecutive embryo recoveries. These were performed 56 days apart. The superovulation begun on 8th day of oestrus synchronization and all goats received 250 UI of porcine FSH splited into eight decreasing dosages at 12 hours intervals. The embryo recovery took place on the 5th or 6th day from the last mating. Fifty-six days after the third recovery of embryos, the animals were sacrified and the genital tract was evaluated. The time spent to recovery the embryos was 21min 32sec; 37min 14sec and 56min 22sec, respectively to T1; T2 and T3 (p<0.01). T3 showed the highest recovery rate of washing solution (83.7%), followed by T2 (72.2%) and T1 (64.3%) (p<0.05). Embryo recovery rate had as mean values, 57.1; 81.1 and 27.3%, respectivily to T1, T2 and T3. The variation ranged between 0 and 100%. Several factors affected the embryo recovery rate, such as the presence of demised corpora lutea, the ovulation rate and the presence of adhesions in the genital tract. Nonetheless, embryo recevery rate was acceptable in all treatments. The T1 caused eversion of endometrium and adherence between the uterine horn and epiploon in one animal, the T2 caused 30, 40 and 60% of endometrium eversion and 10, 10 and 70% adherence on genital tract, respectively, for the first, second and third embryo recovery. The T3 caused 80% of adherence on genital tract after the first embryo recovery and 100% after the second. It is possible to use the methods T1 and T2 to recovery embryos in goats.

Caprine lentivirus in sheep milk and semen

Abstract: With the objective of detecting the presence of caprine lentivirus (CLV) in ewe milk and in ram semen, ten matrixes and four reproducers experimentally infected with CLV were used. Samples of ewe milk were collected during the four months of lactation, five collections per animal, totaling 50 samples. Regarding the rams, eight semen collections were made per animal, during one year of experimentation, totaling 32 samples. The milk and semen samples were submitted to DNA extraction and the nested polymerase chain reaction test (nPCR) to detect CLV proviral DNA. Eight (16%) of the milk samples were positive in nPCR originating from two ewes. Only one (3.12%) semen sample was positive. The amplification products were sequenced, and were confirmed to be a CLV genomic sequence. Thus, the presence of CLV proviral DNA in sheep milk and semen was demonstrated, confirming the feasibility of infection between species, and alerting to the risk of spreading infections. [Lentivirus caprino em leite e semen de ovinos]. Resumo: Com o objetivo de detectar a presenca do lentivirus caprino (LVC) no leite de ovelhas e no semen de carneiros, utilizaram-se 10 matrizes e quatro reprodutores infectados experimentalmente com o LVC. Foram coletadas amostras de leite das ovelhas durante os quatro meses de lactacao, ocorrendo cinco coletas por animal, totalizando 50 amostras. Quanto aos carneiros, realizaram-se oito coletas de semen por animal, durante um ano de experimentacao, totalizando 32 amostras. As amostras de leite e de semen foram submetidas a extracao de DNA e a prova de reacao em cadeia da polimerase do tipo nested (nPCR) visando a deteccao de DNA proviral do LVC. Oito (16%) amostras de leite foram positivas na nPCR oriundas de duas ovelhas. Apenas uma (3,12%) amostra de semen apresentou positividade. Produtos da amplificacao foram sequenciados, confirmando-se tratar de sequencia genomica do LVC. Dessa forma, demonstrou-se a presenca do DNA proviral do LVC em leite e semen de ovinos, confirmando a viabilidade da infeccao entre especies e, assim, alertando sobre o risco de que a infeccao seja disseminada.

Duration of passive immunity to small ruminant lentiviruses in lambs

In order to evaluate the passive immunity against small ruminant lentiviruses (SRLV) in lambs, this study was conducted from two experimental groups. The first one (G1) was established by nine lambs subjected to artificial feeding of colostrum of goats positive for SRLV. The second one (G2) was the control group, consisting of ten lambs subjected to suckling of colostrum from their negative mothers. Blood samples were obtained before the first feeding, after 24 hours of birth and at 7, 15, 30, 50, 70, 90 and 120 days of age. The concentrations of total serum protein (TSP), albumin (ALB), globulin (GLOB) and immunoglobulin G (IgG) were determined and antibodies to SRLV were surveyed from the techniques of agar gel immunodiffusion (AGID), enzyme linked immunosorbent assay (Elisa) and immunoblotting (IB). In both groups, the lowest averages of TSP, GLOB and IgG were observed at birth and the highest averages were observed at 24 hours of life, due to absorption of colostral immunoglobulins. For G1, transfer of immunity could also be detected by immunodiagnostic tests. At birth, the animals were seronegative. After 24 hours, all animals were positive in three serological tests. Negative results began to be observed after 15 days of age by the AGID test. As for Elisa testing, all animals remained reagent until 50 days old. Only IB was able to detect anti-SRLV at 70 days. Regarding G2, all animals tested negative in AGID and IB, from birth until 120 days of age. However, false-positive results were observed until day 15 in Elisa, due to nonspecific reactions. These data are consistent with the sensitivity and specificity of serological tests and show that starting at 90 days of age, colostral antibodies to SRLV are no longer detected in the serum of lambs.

Thoracic aortic aneurysm in a buck associated with caseous lymphadenitis

This paper reports the clinical, bacteriological and pathological findings of a thoracic aortic aneurysm in a four-year-old Anglo-Nubian goat buck, related to a framework of visceral caseous lymphadenitis. General clinical examination showed heart rate of 75 beats per minute, respiratory rate of 20 movements per minute and ruminal movements of four movements per minute. Superficial lymph nodes were normal upon palpation. Rectal temperature was slightly high (40.5°C). Blood test showed an intense leukocytosis (54,000/µL), characterized by strong neutrophil shift to the left. At necropsy, a large blood clot was detected in the thoracic cavity. The thickening of the myocardium and dilatation of the aorta in the thoracic portion, presenting a saculiform format was also observed. A large number of abscesses were disseminated in the media and intima layers of aorta. The aorta lumen obstruction by arterial plaques consisting of inflammatory infiltrate, predominantly neutrophilic was also detected. Abscesses were found spread in turbinate, rumen, reticulum, kidneys, liver, spleen, testicles and aorta wall. The microbiological exam of exudate confirmed Corynebacterium pseudotuberculosis as the causal agent.

Influência da soropositividade ao vírus da artrite encefalite caprina no hemograma de cabras em lactação

The objective was to study the influence of caprine arthritis encephalitis in the blood count of goats during lactation. To this end, 38 F1 Anglo-Nubian x Saanen reproductive female goats, aged 14 to 38 months and body condition score between 2 and 3, were first tested by AGID and Western Blot, and divided into either a seropositive group (n = 19) or a seronegative group (n = 19). The groups were kept in separate paddocks, without physical contact, receiving the same nutritional and health management. The blood of these animals was collected by jugular venipuncture, monthly, from the partum period until the end of lactation, and their erythrogram and leukogram values were determined. At given moments, the seropositive animals showed morphological changes in red blood cells, with significant decrease in MCV, coupled with a significant increase of MCH and MCHC. There was no significant difference between the groups in the values of red blood cells and packed cell volume. The absolute values of monocytes and eosinophils of the seropositive animals, at certain moments, were significantly higher than the seronegative ones and above the normal range for goats.

Infeciosas diseases of small ruminants: economic epidemiologia, impacts, prevention and control: a revision

In this bibliographic review, it is intended to show, on a simple and systematic form, the possible diseases transmitted through meat, milk and other derivates of “caprinos” and “ovinos”. In “caprinos” and “ovinos”, the possible diseases transmitted through meat, milk and other derivates are divided in this present work by two different groups: those occurred by ingesting food named food intoxications, and the “zoonoses”, which bring human and public health risks. The diseases that have a “microbiana” origin, are transmitted through the ingestion of food infected by bacterias such as Escherichia coli, Salmonella sp, Campylobacter sp, Clostridium sp, Staphylococcus sp. On the zoonoses group, it’s possible to talk about “Listeriose, a Brucelose, a Tuberculose, a Toxoplasmose, a Raiva, a Leptospirose e a Febre Aftosa”