Diarrhea in cats infected with Cystoisospora spp. - evaluation of the neutrophil-to-lymphocyte ratio and monocyte-to-lymphocyte ratio

Background: Cystoisospora spp. is considered a potential pathogen to cause diarrhoea in cats, and the disease occurs clinically, especially in young, weak and immunocompromised cats. The present study aimed to evaluate the clinical utility and diagnostic roles of neutrophil-to-lymphocyte ratio (NLR) and monocyte-to-lymphocyte (MLR) ratios in diarrheal cats infected with Cystoisospora spp. Materials, Methods & Results: Twenty-five cats with diarrhea due to infection caused by Cystoisospora spp. and 15 healthy cats were included in this study. The anamnesis, clinical and laboratory findings of all cats were recorded. A clinical severity score was generated for diarrheal cats, which included activity, appetite, presence of vomit, and faecal consistency. Also, cats infected with Cystoisospora spp. were classified as systemic inflammatory response syndrome (SIRS) positive (+) and SIRS (-) group according to the presence of at least 3 of the SIRS criteria. The Cystoisospora spp. oocysts were diagnosed by direct smear and simple faecal flotation. Cats found positive for other parasitic and protozoal agents by faeces examination were excluded from the study. Complete blood counts (CBC) were performed with an automated blood cell counter for all cats. Blood smears were prepared (with May-Grunwald Giemsa) from samples, and the CBC results were confirmed microscopically (obj.100x). White blood cell (WBC), neutrophil, lymphocyte and monocyte counts were recorded directly from the CBC. The neutrophil-to-lymphocyte ratio was calculated as the ratio of the absolute neutrophil count to the absolute lymphocyte count. The monocyte-to-lymphocyte was calculated as the absolute monocyte count ratio to the absolute lymphocyte count. White blood cell (P = 0.015), neutrophil (P = 0.001), monocyte (P = 0.033), NLR (P = 0.000) and MLR (P = 0.042) of diarrheal cats infected with Cystoisospora spp. were significantly higher than in the healthy group. There was no significant difference between groups in the lymphocyte counts (P > 0.05). SIRS (+) group had statistically significantly higher WBC (P = 0.014), neutrophil counts (P = 0.000), NLR (P = 0.000) and MLR (P = 0.037) than the healthy group. Also, neutrophil counts and NLR of the SIRS (+) group were statistically significantly higher than the SIRS (-) group (P = 0.032; P = 0.001, respectively). However, there was no significant difference SIRS (+), SIRS (-), and healthy groups regarding lymphocyte and monocyte counts (P > 0.05). The clinical severity of the disease positively correlated with NLR and MLR. Also, the best cut-off value of NLR to predict SIRS was > 1.67, with 92.86% sensitivity and 84.62% specificity. MLR was not a useful predictor for SIRS. Discussion: NLR and MLR are parameters derived from complete blood count. In recent years, they have been used in human and veterinary medicine as a biomarker in the diagnosis, prognosis and treatment follow-up of various diseases and provide valuable information about inflammation. In cats, there are limited studies on NLR, and to our knowledge, there are no studies on MLR. The current study suggests that NLR and MLR may be used to determine the severity of the inflammatory condition in diarrheal cats infected with Cystoisospora spp., and NLR is a useful marker for predicting SIRS status in these cats. In addition, this study also will contribute to larger-scale studies in the future.

New findings of intestinal alkaline phosphatase: effects on intestinal and organ health of piglets challenged with ETEC F4 (K88)

The aim of this study was to assess the addition of intestinal alkaline phosphatase (IAP) to diets on the count of bacterial populations, pH of digestive organ contents, histopathological description, proinflammatory markers, hepatic glycogen reserve, and diarrhoea incidence of piglets challenged with Escherichia coli. Sixty-four crossbred piglets (7.16±0.28 kg body weight, 25-days-old) were assigned to four treatments in a completely randomised block design: negative control (NC), NC + antibiotic (ANT), NC + 15 mg IAP, or NC + 30 mg IAP kg−1 of diet, eight replications of two piglets per experimental unit. All piglets were orally challenged with 6 mL of a solution containing enterotoxigenic Escherichia coli K88 at 106 CFU mL−1 at 15 days of experimentation. The study lasted for 19 days. At the end of the experimental period, the piglets were slaughtered (six animals per treatment). Enterobacteriaceae in caecum and colon was lower in piglets on 30 mg IAP than with ANT and NC, ANT or 15 mg IAP, respectively. Enterobacteriaceae adhered to the mesenteric lymph nodes (MLN) was greater in piglets fed ANT than the other treatments. Lactic acid bacteria (LAB) count in caecum was greater in piglets fed NC and ANT. In MLN, LAB count was greater in ANT and 30 mg IAP-fed piglets compared with 15 mg IAP. Piglets in 30 mg IAP in diet showed a tendency for lowering tissue necrosis compared with NC or ANT. Piglets fed 30 mg IAP showed a reduction in diarrhoea incidence in the pre- and post-challenge compared with 15 mg IAP and all other treatments, respectively. Based on the criteria, addition of 30 mg IAP to diet inhibits Enterobacteriaceae population and suggests a potential effect in mitigating intestinal injuries, as observed in piglets in the NC for some of the parameters investigated.(AU)

Dietary fiber and zinc additives on performance and intestinal health of Escherichia coli challenged piglets

This study aimed to evaluate the effects of zinc oxide and a low level of encapsulated zinc oxide, with or without dietary fiber, on the performance and intestinal health of weaned piglets. A total of 112 piglets were used, divided into four treatment groups: basal diet with zinc oxide (ZnO); basal diet with zinc oxide and dietary fiber (ZnO+DF); basal diet with encapsulated low zinc oxide (LZnOE); and basal diet with LZnOE and DF (LZnOE+DF). Piglets were challenged with E . coli K88 + , weighed weekly and the fecal score was evaluated daily. One pig per pen was slaughtered to evaluate the production of volatile fatty acids, intestinal microbial populations, intestinal morphology, and digestibility. The encapsulated zinc resulted in lower body weight and average daily gain, but, when associated with dietary fiber, had similar results to zinc oxide. Until 49 days of age, zinc oxide reduced diarrhoea ( p 0.05). At 63 days of age the piglets subjected to LZnOE+DF treatment had lower diarrhoea than the pigs subjected to LZnOE treatment but higher than the ZnO and the ZnO+DF groups. Final weight and incidence of diarrhoea of pigs receiving LZnOE was worse than ZnO. LZnOE associated with DF provided the same final weight of piglets from the ZnO group.

Dietary fiber and zinc additives on performance and intestinal health of Escherichia coli challenged piglets

This study aimed to evaluate the effects of zinc oxide and a low level of encapsulated zinc oxide, with or without dietary fiber, on the performance and intestinal health of weaned piglets. A total of 112 piglets were used, divided into four treatment groups: basal diet with zinc oxide (ZnO); basal diet with zinc oxide and dietary fiber (ZnO+DF); basal diet with encapsulated low zinc oxide (LZnOE); and basal diet with LZnOE and DF (LZnOE+DF). Piglets were challenged with E . coli K88 + , weighed weekly and the fecal score was evaluated daily. One pig per pen was slaughtered to evaluate the production of volatile fatty acids, intestinal microbial populations, intestinal morphology, and digestibility. The encapsulated zinc resulted in lower body weight and average daily gain, but, when associated with dietary fiber, had similar results to zinc oxide. Until 49 days of age, zinc oxide reduced diarrhoea ( p 0.05). At 63 days of age the piglets subjected to LZnOE+DF treatment had lower diarrhoea than the pigs subjected to LZnOE treatment but higher than the ZnO and the ZnO+DF groups. Final weight and incidence of diarrhoea of pigs receiving LZnOE was worse than ZnO. LZnOE associated with DF provided the same final weight of piglets from the ZnO group.(AU)

Additional effect of coated sodium butyrate 30% in the piglets feeding during the nursery phase / Efeito adicional do butirato de sódio 30% revestido na alimentação de leitões durante a fase de creche

In this study, the objective was to assess the growth performance, rectal microbiological profile, serum and immunological standards, and incidence of diarrhoea in piglets in the nursery phase fed diets with and without the addition of coated sodium butyrate 30% (SB). A total of 80 crossbred piglets (40 entire males and 40 females), with an average initial body weight of 9.47 ± 1.03 kg, were distributed in a randomised block design, with four treatments repeated twice in the two blocks and five piglets of the same gender per experimental unit. The treatments comprised a combination of two feeds, formulated based on corn and soybean meal with and without 0.1% SB inclusion as an additional product, and both genders, fed from of the pre-starter I diet. SB did not affect (p > 0.05) the growth performance and rectal microbiological profile of the piglets, but efficiently (p = 0.05) controlled the incidence of diarrhoea. The average values of the characteristics measured in the blood of the piglets were within the normal physiological parameters of the species. The inclusion of SB in commercial feed did not alter the growth performance and rectal microbiological profile of nursery piglets under low sanitary challenge conditions. However, the organic acid efficiently reduced gastrointestinal disorders that cause diarrhoea, with diarrhoea incidence rates of 2.68 and 4.46% for piglets fed with and without the inclusion of the additional organic acid, respectively.(AU)

Neste estudo, o objetivo foi o de avaliar o desempenho zootécnico, o perfil microbiológico retal, padrões imunológicos e no soro, e a incidência de diarreia em leitões na fase de creche alimentados com dietas adicionadas ou não do butirato de sódio 30% revestido (BS). Um total de 80 leitões mestiços (40 machos inteiros e 40 fêmeas), com um peso corporal médio inicial de 9,47 ± 1,03 kg, distribuídos em um delineamento experimental de blocos casualizados, com quatro tratamentos repetidos duas vezes nos dois blocos e cinco suínos de mesmo sexo por unidade experimental. Os tratamentos foram constituídos pela combinação de duas rações formuladas à base de milho e farelo de soja, com e sem inclusão de 0,1% de BS como produto adicional, e dois sexos, alimentados a partir da dieta pré-inicial I. Não houve efeito (p > 0,05) do BS no desempenho zootécnico e no perfil microbiológico retal dos leitões, mas houve eficiência do BS (p = 0,05) no controle da incidência de diarreia. Os valores médios das características mensuradas no sangue dos leitões estão dentro dos parâmetros fisiológicos normais da espécie. A inclusão de BS em rações comerciais não altera o desempenho zootécnico e o perfil microbiológico retal de leitões na fase de creche sob condições de baixo desafio sanitário. Entretanto, o ácido orgânico é eficiente na redução de distúrbios gastrointestinais causadores de diarreia, com proporções de incidência de diarreia de 2,68% e 4,46% para leitões alimentados com e sem inclusão do ácido orgânico adicional, respectivamente.(AU)

Analysis of risk factors and clinical manifestations associated with Clostridium difficile disease in Serbian hospitalized patients

Clostridium difficile is the leading cause of infectious diarrhoea in hospitalized patients. The aim of this study was to determine the risk factors important for the development of hospital-acquired Clostridium difficile-associated disease and clinical manifestations of Clostridium difficile-associated disease. The clinical trial group included 37 hospitalized patients who were selected according to the inclusion criteria. A control group of 74 hospitalized patients was individually matched with cases based on hospital, age (within 4 years), sex and month of admission.Clostridium difficile-associated disease most commonly manifested as diarrhoea (56.76%) and colitis (32%), while in 8.11% of patients, it was diagnosed as pseudomembranous colitis, and in one patient, it was diagnosed as fulminant colitis. Statistically significant associations (p 0.05) were found with the presence of chronic renal failure, chronic obstructive pulmonary disease, cerebrovascular accident (stroke) and haemodialysis. In this study, it was confirmed that all the groups of antibiotics, except for tetracycline and trimethoprim-sulfamethoxazole, were statistically significant risk factors for Clostridium difficile-associated disease (p 0.05). However, it was difficult to determine the individual role of antibiotics in the development of Clostridium difficile-associated disease.[...](AU)

Bovine coronavirus (BoCV) infection in calves with diarrhoea and their dams

Background: Bovine coronavirus (BoCV) is common with high seroprevalence in dairy cattle. It is reported in many countries. Also, BoCV causes diarrhea in dairy calves. The transmission of BoCV is the fecal-oral/aerosol-nasal routes. Feces from clinical cases or clinically normal dairy cattle are source of infection, also contamination of feed and water. The purpose of the current study was to investigate BoCV infection in diarrheic calves (age and sex) and their dams (age). For this reason, the serological and virological methods were used. Haematological parameters of the calves and their dams were compared using the statistical methods.Materials, Methods & Results: In this study, following clinical examination of 3500 cattle and their calves from 25 number of dairy farms 184 calves with diarrhoea and their dams (183) (2 - 6 age) were sampled for BoCV presence by enzyme linked immunosorbent assay (ELISA). Additionally, all blood samples were examined by hematological methods. 172 (93.99%) cows and 172 (93.99%) calves were found antibodies (Ab) positive (+). The high levels of Ab for BoCV were detected as 36.05 % in dams 6 years and older ages. In the calves, Ab to BoCV were found at the highest level (25.26%) in the female calves 5 - 6 months ages. BoCV antigen (Ag) was detected in only faecal sample of a (0.54%) calf. When the haematological parameters were compared [...](AU)

Bovine coronavirus (BoCV) infection in calves with diarrhoea and their dams

Background: Bovine coronavirus (BoCV) is common with high seroprevalence in dairy cattle. It is reported in many countries. Also, BoCV causes diarrhea in dairy calves. The transmission of BoCV is the fecal-oral/aerosol-nasal routes. Feces from clinical cases or clinically normal dairy cattle are source of infection, also contamination of feed and water. The purpose of the current study was to investigate BoCV infection in diarrheic calves (age and sex) and their dams (age). For this reason, the serological and virological methods were used. Haematological parameters of the calves and their dams were compared using the statistical methods.Materials, Methods & Results: In this study, following clinical examination of 3500 cattle and their calves from 25 number of dairy farms 184 calves with diarrhoea and their dams (183) (2 - 6 age) were sampled for BoCV presence by enzyme linked immunosorbent assay (ELISA). Additionally, all blood samples were examined by hematological methods. 172 (93.99%) cows and 172 (93.99%) calves were found antibodies (Ab) positive (+). The high levels of Ab for BoCV were detected as 36.05 % in dams 6 years and older ages. In the calves, Ab to BoCV were found at the highest level (25.26%) in the female calves 5 - 6 months ages. BoCV antigen (Ag) was detected in only faecal sample of a (0.54%) calf. When the haematological parameters were compared [...]

Prevalencia de anticuerpos para el Vírus de la diarreia viral bovina, Herpesvírus bovino Tipo 1 y Vírus de la leucosis enzoótica bovina en búfalos de rio de la Isla de Marajó / Prevalence of Bovine Viral Diarrhoea Virus, Bovine Herpesvirus 1 and Bovine Leucosis Virus antibodies in river buffaloes from Marajó Island / Prevalência de anticorpos para o Vírus da diarreia viral bovina, Herpesvírus bovino Tipo 1 e Vírus da leucose enzoótica bovina em búfalos de água da Ilha de Marajó

El objetivo de este estudio fue determinar la prevalencia de anticuerpos en una manada de búfalos de río en la Isla de Marajó en busca de virus: Virus de la Diarrea Viral Bovina, Bovino Herpesvirus tipo 1 del Virus y la leucosis bovina enzoótica. La técnica de neutralización se utilizó para detectar anticuerpos relacionados con el virus de la diarrea viral bovina y virus del herpes bovino tipo 1 y la prueba de inmunodifusión en gel de agar para la detección de anticuerpos contra el virus de la leucosis bovina enzoótica. La prevalencia de anticuerpos contra el virus del herpes bovino tipo 1 en los búfalos está muy cerca de la prevalencia observada en el ganado bovino. La prevalencia de anticuerpos viral bovina virus de la diarrea mostró muy por debajo de la media en comparación con la prevalencia de anticuerpos en el ganado bovino y búfalos, también creado en el Brasil. La prevalencia del virus de la leucosis bovina enzoótica mostró, sin embargo, cerca del rango observado en búfalos en Brasil y en todo el mundo, que divergen de la prevalencia observada en los anticuerpos de ganado muy bajo, probablemente mostrando una resistencia natural al virus de este tipo, este factor debe ser mejor dilucidado.(AU)

The aim of this study was to determine the antibodies prevalence of Bovine viral diarrhea virus, Bovine herpesvirus 1 and Bovine leukemia virus in a herd of river buffaloes from the Marajó Island. The serum neutralization technique was used for the detection of Bovine viral diarrhea virus and Bovine herpesvirus 1 antibodies. The agar gel immunodiffusion (AGID) was used for the detection of Bovine leukemia virus antibodies. The prevalence of antibody to the Bovine herpesvirus 1 in buffaloes is very similar to the observed in cattle. The Bovine viral diarrhea virus showed a below average prevalence whether compared to the antibodies prevalence in cattle and buffaloes created in other parts of Brazil. The Bovine leukemia virus showed a very low prevalence of antibodies, however, these parameters were similar to others observed in buffaloes in Brazil and worldwide. These parameters differ from the prevalence observed in cattle, which probably suggests a natural resistance from this specie to the virus. This fact, however, needs to be further elucidated.(AU)

O objetivo do presente estudo foi determinar a prevalência de anticorpos em um rebanho de búfalos de rio da Ilha de Marajó para os vírus: Vírus da diarreia viral bovina, Herpesvírus bovino tipo 1 e Vírus da leucose enzoótica bovina. A técnica de soroneutralização foi utilizada para detecção de anticorpos referentes aos Vírus da diarreia viral bovina e Herpesvírus bovino tipo 1 e o teste de imunodifusão em gel de ágar para detecção de anticorpos para o Vírus da leucose enzoótica bovina. A prevalência de anticorpos para o Herpesvírus bovino tipo 1 em bubalinos está muito próxima da prevalência observada em bovinos. O Vírus da diarreia viral bovina apresentou prevalência de anticorpos muito abaixo da média quando comparada com prevalência de anticorpos em bovinos e bubalinos, inclusive criados no Brasil. O Vírus da leucose enzoótica bovina mostrou prevalência de anticorpos muito baixa, porém, próxima dos parâmetros observados em búfalos no Brasil e no mundo, que divergem da prevalência observada em bovinos, mostrando provavelmente uma resistência natural dessa espécie ao vírus, fator esse que precisa ser mais bem elucidado.(AU)

Prevalencia de anticuerpos para el Vírus de la diarreia viral bovina, Herpesvírus bovino Tipo 1 y Vírus de la leucosis enzoótica bovina en búfalos de rio de la Isla de Marajó / Prevalence of Bovine Viral Diarrhoea Virus, Bovine Herpesvirus 1 and Bovine Leucosis Virus antibodies in river buffaloes from Marajó Island / Prevalência de anticorpos para o Vírus da diarreia viral bovina, Herpesvírus bovino Tipo 1 e Vírus da leucose enzoótica bovina em búfalos de água da Ilha de Marajó

El objetivo de este estudio fue determinar la prevalencia de anticuerpos en una manada de búfalos de río en la Isla de Marajó en busca de virus: Virus de la Diarrea Viral Bovina, Bovino Herpesvirus tipo 1 del Virus y la leucosis bovina enzoótica. La técnica de neutralización se utilizó para detectar anticuerpos relacionados con el virus de la diarrea viral bovina y virus del herpes bovino tipo 1 y la prueba de inmunodifusión en gel de agar para la detección de anticuerpos contra el virus de la leucosis bovina enzoótica. La prevalencia de anticuerpos contra el virus del herpes bovino tipo 1 en los búfalos está muy cerca de la prevalencia observada en el ganado bovino. La prevalencia de anticuerpos viral bovina virus de la diarrea mostró muy por debajo de la media en comparación con la prevalencia de anticuerpos en el ganado bovino y búfalos, también creado en el Brasil. La prevalencia del virus de la leucosis bovina enzoótica mostró, sin embargo, cerca del rango observado en búfalos en Brasil y en todo el mundo, que divergen de la prevalencia observada en los anticuerpos de ganado muy bajo, probablemente mostrando una resistencia natural al virus de este tipo, este factor debe ser mejor dilucidado.

The aim of this study was to determine the antibodies prevalence of Bovine viral diarrhea virus, Bovine herpesvirus 1 and Bovine leukemia virus in a herd of river buffaloes from the Marajó Island. The serum neutralization technique was used for the detection of Bovine viral diarrhea virus and Bovine herpesvirus 1 antibodies. The agar gel immunodiffusion (AGID) was used for the detection of Bovine leukemia virus antibodies. The prevalence of antibody to the Bovine herpesvirus 1 in buffaloes is very similar to the observed in cattle. The Bovine viral diarrhea virus showed a below average prevalence whether compared to the antibodies prevalence in cattle and buffaloes created in other parts of Brazil. The Bovine leukemia virus showed a very low prevalence of antibodies, however, these parameters were similar to others observed in buffaloes in Brazil and worldwide. These parameters differ from the prevalence observed in cattle, which probably suggests a natural resistance from this specie to the virus. This fact, however, needs to be further elucidated.

O objetivo do presente estudo foi determinar a prevalência de anticorpos em um rebanho de búfalos de rio da Ilha de Marajó para os vírus: Vírus da diarreia viral bovina, Herpesvírus bovino tipo 1 e Vírus da leucose enzoótica bovina. A técnica de soroneutralização foi utilizada para detecção de anticorpos referentes aos Vírus da diarreia viral bovina e Herpesvírus bovino tipo 1 e o teste de imunodifusão em gel de ágar para detecção de anticorpos para o Vírus da leucose enzoótica bovina. A prevalência de anticorpos para o Herpesvírus bovino tipo 1 em bubalinos está muito próxima da prevalência observada em bovinos. O Vírus da diarreia viral bovina apresentou prevalência de anticorpos muito abaixo da média quando comparada com prevalência de anticorpos em bovinos e bubalinos, inclusive criados no Brasil. O Vírus da leucose enzoótica bovina mostrou prevalência de anticorpos muito baixa, porém, próxima dos parâmetros observados em búfalos no Brasil e no mundo, que divergem da prevalência observada em bovinos, mostrando provavelmente uma resistência natural dessa espécie ao vírus, fator esse que precisa ser mais bem elucidado.

Adherence and virulence genes of Escherichia coli from children diarrhoea in the Brazilian Amazon

The bacterial pathogen most commonly associated with endemic forms of childhood diarrhoea is Escherichia coli. Studies of epidemiological characteristics of HEp-2 cell-adherent E. coli in diarrhoeal disease are required, particularly in developing countries. The aim of this study was evaluate the presence and significance of adherent Escherichia coli from diarrhoeal disease in children. The prevalence of LA, AA, and DA adherence patterns were determined in HEp-2 cells, the presence of virulence genes and the presence of the O serogroups in samples obtained from 470 children with acute diarrhoea and 407 controls in Porto Velho, Rondônia, Brazil. E. coli isolates were identified by PCR specific for groups of adherent E. coli. Out of 1,156 isolates obtained, 128 (11.0%) were positive for eae genes corresponding to EPEC, however only 38 (29.6%) of these amplified bfpA gene. EAEC were isolated from 164 (14.1%) samples; of those 41(25%), 32 (19%) and 16 (9.7%) amplified eagg, aggA or aafA genes, respectively and aggA was significantly associated with diarrhoea (P = 0.00006). DAEC identified by their adhesion pattern and there were few isolates. In conclusion, EAEC was the main cause of diarrhoea in children, especially when the aggA gene was present, followed by EPEC and with a negligible presence of DAEC.(AU)

Colicin type 7 produced by majority of Shigella sonnei isolated from Thai patients with diarrhoea

Thirty one out of 153 strains of Shigella sonnei isolated from Thai patients with diarrhoea showed antibacterial activity against S. sonnei by agar well diffusion method. All of them harbor plasmids with the genetic determination of colicin type 7 (Js) gene but without colicin E and colicin U gene. The PCR product obtained from strain 35/44 was shown to be the gene for colicin type 7 lytic protein (cja). The partially purified bacteriocin (PPB) containing colicin type 7 of strain 35/44 was prepared and used for characterization. The antibacterial activity of PPB against a total of 17 selected Gram-positive and Gram-negative bacteria was tested. It was found that PPB of strain 35/44 was active against E. coli O157, S. sonnei and S. boydii. The sensitivity of PPB from this strain to proteinase K, trypsin and -chymotrypsin suggests the proteinaceous nature of these antimicrobial substances. Therefore, this isolated bacterium can be regarded as bacteriocin producing bacteria. The bacteriocin produced by this isolated S. sonnei was heat stable as evidenced by its ability to maintain the activity at 80 °C for 60 min. In addition, it was stable within a wide range of pH (3-9). The molecular weight of colicin type 7 from isolated S. sonnei strain 35/44 analyzed by SDS-PAGE was 54.4 kDa composing of at least five subunits. It is to our knowledge, the first report of Thai patients with diarrhoea that S. sonnei isolated from them contained colicin type 7.(AU)

Genetic characterization of bovine viral diarrhoea virus detected in persistently infected cattle in Southern Minas Gerais, Brazil / Caracterização genética do vírus da diarreia bovina detectada em bovinos do Sul de Minas Gerais, Brasil, persistentemente infectados

Isolates of bovine viral diarrhoea virus (BVDV) detected in serum samples of two persistently infected animals (PI) identified in a herd located in the southern state of Minas Gerais, Brazil, underwent genetic characterization trough partial nucleotide sequencing and analysis of the 5 Untranslated Region (5UTR) of the viral genome. The isolates were characterized as belonging to genotype BVDV-1, subgenotype BVDV-1b. The results of this study suggest BVDV-1b as an agent of importance in the occurrence of bovine viral diarrhoea (BVD) in the herds of the region. Moreover, the genotypic characterization of isolates of BVDV helps to better understand the epidemiology of the disease, as the genetic variability of BVDV interferes in the serological tests and has implications for the use of vaccines, whose majority is produced only with reference strains of BVDV. Therefore, the investigation on the genetic diversity of BVDV existing in Brazil is required for the improvement of the disease prevention and control measures.(AU)

Isolados do vírus da diarreia viral bovina (BVDV) detectados em amostras de soro sanguíneo de dois animais persistentemente infectados (PI), identificados num rebanho bovino localizado na região sul do Estado de Minas Gerais, Brasil, foram submetidos à caracterização genética através do sequenciamento parcial de nucleotídeos da região 5UTR do genoma viral. Os isolados foram caracterizados como pertencentes ao genótipo BVDV-1, subgenótipo BVDV-1b. Os resultados do presente estudo sugerem o BVDV-1b como um agente de importância na ocorrência da diarreia viral bovina (BVD) nos rebanhos da região. Ademais, a caracterização genotípica dos isolados do BVDV contribui para a melhor compreensão da epidemiologia da enfermidade, pois a variabilidade genética do BVDV interfere nos testes sorológicos e também possui implicações na utilização de vacinas, cuja maioria é produzida apenas com estirpes de referência do BVDV, requerendo, portanto, investigações sobre a diversidade genética do BVDV existente no Brasil para o aprimoramento das medidas de prevenção e controle da enfermidade no país.(AU)

Genetic characterization of bovine viral diarrhoea virus detected in persistently infected cattle in Southern Minas Gerais, Brazil / Caracaterização genética do vírus da diarréia viral bovina detectado em bovinos persistentemente infectados na região sul do estado de Minas Gerais, Brasil

Isolates of bovine viral diarrhoea virus (BVDV) detected in serum samples of two persistently infected animals (PI) identified in a herd located in the southern state of Minas Gerais, Brazil, underwent genetic characterization trough partial nucleotide sequencing and analysis of the 5 Untranslated Region (5UTR) of the viral genome. The isolates were characterized as belonging to genotype BVDV-1, subgenotype BVDV-1b. The results of this study suggest BVDV-1b as an agent of importance in the occurrence of bovine viral diarrhoea (BVD) in the herds of the region. Moreover, the genotypic characterization of isolates of BVDV helps to better understand the epidemiology of the disease, as the genetic variability of BVDV interferes in the serological tests and has implications for the use of vaccines, whose majority is produced only with reference strains of BVDV. Therefore, the investigation on the genetic diversity of BVDV existing in Brazil is required for the improvement of the disease prevention and control measures.(AU)

Isolados do vírus da diarreia viral bovina (BVDV) detectados em amostras de soro sanguíneo de dois animais persistentemente infectados (PI), identificados num rebanho bovino localizado na região sul do Estado de Minas Gerais, Brasil, foram submetidos à caracterização genética através do sequenciamento parcial de nucleotídeos da região 5UTR do genoma viral. Os isolados foram caracterizados como pertencentes ao genótipo BVDV-1, subgenótipo BVDV-1b. Os resultados do presente estudo sugerem o BVDV-1b como um agente de importância na ocorrência da diarreia viral bovina (BVD) nos rebanhos da região. Ademais, a caracterização genotípica dos isolados do BVDV contribui para a melhor compreensão da epidemiologia da enfermidade, pois a variabilidade genética do BVDV interfere nos testes sorológicos e também possui implicações na utilização de vacinas, cuja maioria é produzida apenas com estirpes de referência do BVDV, requerendo, portanto, investigações sobre a diversidade genética do BVDV existente no Brasil para o aprimoramento das medidas de prevenção e controle da enfermidade no país.(AU)

Desenvolvimento PCR em Tempo-Real em sistema TaqMan® para detecção de rotavírus em amostras clínicas de bovinos

A diarréia neonatal é o principal problema sanitário que acomete os bezerros nas primeiras semanas de vida, causando grandes prejuízos devido à morbidade, mortalidade, custos com tratamento e atraso no desenvolvimento. Os rotavírus são os mais importantes agentes virais causadores de gastroenterites em crianças e diarreia em diferentes espécies animais. Além do seu impacto econômico na produção animal, os bovinos podem atuar como reservatórios da diversidade genética e antigênica para humanos. Assim, o diagnóstico deste agente é fundamental para o desenvolvimento de medidas profiláticas mais especificas visando o seu controle. O objetivo deste trabalho foi desenvolver um método de PCR em Tempo Real para a detecção de rotavírus bovinos em sistema Taqman® tendo como alvo, a proteína nãoestrutural 5 (NSP5). Para isso, 113 amostras de fezes bovinas foram coletadas de reabanhos do estado de São Paulo, e previamente testadas por PCR convencional.Para a padronização da PCR em Tempo Real , a estirpe padrão foi clonada, gerando um plasmídeo com 3x1010 cópias/reação. Como controle exógeno foi utilizado -actina. O limite de detecção foi determinado por diluições seriadas na base 10, detectando até 6x101 cópias/l. A curva padrão da PCR em Temo Real para a detecção do segmento codificador da proteína NSP5 teve como resultados, uma eficiência de 100,47%; com slope igual a -3,18 e R2 de 1,0. De um total de 113 amostras testadas pela PCR convencional 63 delas (55,7%) foram consideradas positivas para rotavírus. Dessas mesmas amostras testadas, 5 não amplificaram para -actina e não foram incluídas nas análises posteriores. Para a PCR em Tempo Real o limite de detecção foi considerado o valor de 6x101 cópias/reação, sendo definido o ponto de corte o ciclo (Ct) de número 36 para o teste com a amostra viral a partir do DNA ligado em vetor plasmidial. Considerando-se o ponto de corte de 60 (6x101) cópias/reação, das 108 amostras testadas, 63 (58,3%) foram consideradas positivas ao teste. O valor de concordância obtido através do teste Kappa, a um intervalo de confiança de 95%, a partir dos resultados gerados entre os testes de PCR convencional e PCR em foi de 0.279 (baixa concordância). Os resultados obtidos nesse estudo demonstrou que o teste foi eficiente podendo ser utilizado para um diagnóstico rápido e eficiente do rotavirus do grupo A, aumentando assim o repertório dos testes já estabelecidos.

Neonatal diarrhea is the main health problem affecting the calves in the first weeks of life, causing major losses due to morbidity, mortality, treatment costs and delayed development. Rotaviruses are the most important causative agents of viral gastroenteritis in children and in different animal species. In addition to its economic impact on livestock, cattle can act as reservoirs for genetic and antigenic diversity for human samples. Therefore, the diagnosis of this agent is critical to the development of more specific preventive measures for their control. The objective of this work is to develop a method of PCR for rotavirus detection in cattle using TaqMan® system targeting the 5 nonstructural protein (NSP5). For this, 113 samples of cattle feces were collected from farms of São Paulo, and previously tested by convencional PCR. To PCR standardization, the standard sample was cloned, generating plasmid 3x1010 copies/reaction. The -actin was usede as exogenous control. The limit of detection was determined by serial dilutions, to detect 6 x101 copies/l. The standard curve of PCR to encoding segment detection NSP5 protein had as a result, an efficiency of 108.5%; with slope equal to -3.18 and R2 equal 1.. From a total of 113 samples tested by conventional PCR 63 (55.7%) were positive for rotavirus. From these samples 5 not amplifyed for -actin gene and were not included in subsequent analyzes. For detection limit of Real-Time PCR was considered the amount of 6x101 copies / reaction, the cut-off being defined cycle (Ct) number 36 to the test with a viral sample from the DNA ligated into plasmid vector. Considering the cut-off 60 (6x101) copies/reaction of the 108 samples tested, 63 (58.3%) were positive to the test. The correlation value obtained by the Kappa test, at a 95% confidence interval, based on results generated between the conventional and PCR testing PCR was 0.279 (low agreement). The results of this study demonstrated that the probe can be efficiently used for a fast and efficient diagnosis of rotavirus of group A, thereby enhancing the repertoire of the established tests.

Ocorrência de animais persistentemente infectados pelo vírus da diarréia viral bovina em rebanhos bovinos nos Estados de Minas Gerais e São Paulo / Occurrence of persistently infected animals with bovine viral diarrhoea virus in cattle herds from the States of Minas Gerais and São Paulo, Brazil

A pesquisa de animais persistentemente infectados (PI) pelo vírus da diarréia viral bovina (BVDV) foi realizada em 26 rebanhos bovinos, não vacinados contra o BVDV, localizados nos Estados de Minas Gerais e São Paulo, Brasil. Utilizando uma estratégia de amostragem, de cada rebanho foram obtidas cinco amostras de sangue de bezerros, entre 6 e 12 meses de idade, e os soros sanguíneos foram submetidos ao teste de virusneutralização (VN) para o BVDV-1 e o BVDV-2. Os rebanhos que apresentaram pelo menos três das cinco amostras reagentes a um dos genótipos do BVDV, e com títulos de anticorpos superiores a 128, foram selecionados para a pesquisa de animais PI. Em três rebanhos que apresentaram tal condição, foram colhidas amostras pareadas de sangue de todos os bovinos do rebanho, com intervalo de 30 dias entre as colheitas, e o soro sanguíneo foi submetido ao teste de VN para o BVDV-1 e o BVDV-2. Nas amostras não reagentes a pelo menos um dos genótipos do BVDV e naquelas provenientes de bovinos com menos de seis meses de idade, realizou-se a pesquisa do BVDV pela reação em cadeia da polimerase precedida pela transcrição reversa (RT-PCR). Dos rebanhos analisados, foram detectados dois animais PI a partir de amostras obtidas nas colheitas pareadas provenientes de um rebanho localizado no Estado de Minas Gerais.(AU)

The research on persistently infected (PI) animals with bovine viral diarrhoea virus (BVDV) was conducted in 26 cattle herds, which were not BVDV vaccinated, located in the states of Minas Gerais and São Paulo, Brazil. Using a sampling strategy, five samples of blood were collected from 6 to 12-month-old calves of each herd, and the blood sera were tested by virusneutralization test (VN) to BVDV-1 and BVDV-2. The herds that had at least three out of five samples reacting to one of the genotypes of BVDV and antibody titers greater than 128 were selected to PI animals research. In three of the herds that matched the before-mentioned criteria, paired blood samples were collected from all its individuals considering a collection interval of 30 days. The blood sera of these samples were VN tested against BVDV-1 and BVDV-2. In samples not reacting to at least one of the BVDV genotypes and also in those collected from calves of less than six months of age, virus research was undertaken by reverse transcription polymerase chain reaction (RT-PCR). From the examined herds, two PI animals were detected in paired samples obtained from a herd located in the state of Minas Gerais.(AU)

Absence of diarrhoea in equidae treated with oxytetracycline / Ausência de diarréia em eqüídeos medicados com oxitetraciclina

Evaluation and observation was carried out with 24 equidae put on oxytetracycline hydrochloride medication, a drug incriminated as potential cause of severe colitis with diarrhea manifestations for this species. The animals of this study were natural carriers of affections that required antimicrobial use. During 4 to 19 uninterrupted days intravenous doses of 7, 15 or 20mg/kg were administered, every 24 hours. No side effects were observed during the period of treatment or after it. Thus, at the doses and intervals described above, oxytetracycline showed to be safe for equidae.

Foram avaliados e acompanhados 24 eqüídeos medicados com cloridrato de oxitetraciclina, fármaco incriminado como potencial causador de colite severa com manifestação de diarréia nesta espécie. Os eqüídeos estudados eram portadores naturais de afecções que requeriam o emprego de antimicrobianos. Administraram-se doses de 7, 15 ou 20mg/kg, a cada 24 horas, por via intravenosa, durante 4 a até 19 dias seguidos. Não se observou qualquer efeito colateral aparente durante o período de tratamento e após o seu término. A oxitetraciclina mostrou-se, portanto, segura para os eqüídeos, nas doses e intervalo entre doses utilizados.

Ocurrence of Escherichia Coli, rotavirus, picobirnavirus and Cryptosporidium Parvum in a postweaning diarrhoea focus in swinw / Ocorrência de Escherichia Coli, rotavirus, picobirnavirus e Cryptosporidium Parvum em um foco de diarréia do pós-desmame em suínos

This work describes the etiology of a postweaning diarrhoea focus in a intensive piggery. The results of identification procedures showed the simultaneous occurrence of Escherichia coli, rotavirus, picobirnavirus and Cryptosporidium parvum. In VERO cells culture, was detected the LT (termolabile toxin) production by three E. coli samples and VT (verotoxin) production by five samples.

No presente trabalho é descrita a etiologia de um foco de diarréia em leitões de creche em uma granja de ciclo completo. Os exames laboratoriais mostraram o envolvimento simultâneo de E. coli, rotavirus, picobirnavírus e Cryptosporidium parvum. Em-células VERO três amostras de E. coli produziram LT(toxina termolábil) e cinco VT (verotoxina).

Absence of diarrhoea in equidae treated with oxytetracycline / Ausência de diarréia em eqüídeos medicados com oxitetraciclina

Evaluation and observation was carried out with 24 equidae put on oxytetracycline hydrochloride medication, a drug incriminated as potential cause of severe colitis with diarrhea manifestations for this species. The animals of this study were natural carriers of affections that required antimicrobial use. During 4 to 19 uninterrupted days intravenous doses of 7, 15 or 20mg/kg were administered, every 24 hours. No side effects were observed during the period of treatment or after it. Thus, at the doses and intervals described above, oxytetracycline showed to be safe for equidae.

Foram avaliados e acompanhados 24 eqüídeos medicados com cloridrato de oxitetraciclina, fármaco incriminado como potencial causador de colite severa com manifestação de diarréia nesta espécie. Os eqüídeos estudados eram portadores naturais de afecções que requeriam o emprego de antimicrobianos. Administraram-se doses de 7, 15 ou 20mg/kg, a cada 24 horas, por via intravenosa, durante 4 a até 19 dias seguidos. Não se observou qualquer efeito colateral aparente durante o período de tratamento e após o seu término. A oxitetraciclina mostrou-se, portanto, segura para os eqüídeos, nas doses e intervalo entre doses utilizados.

Ocurrence of Escherichia Coli, rotavirus, picobirnavirus and Cryptosporidium Parvum in a postweaning diarrhoea focus in swinw / Ocorrência de Escherichia Coli, rotavirus, picobirnavirus e Cryptosporidium Parvum em um foco de diarréia do pós-desmame em suínos

This work describes the etiology of a postweaning diarrhoea focus in a intensive piggery. The results of identification procedures showed the simultaneous occurrence of Escherichia coli, rotavirus, picobirnavirus and Cryptosporidium parvum. In VERO cells culture, was detected the LT (termolabile toxin) production by three E. coli samples and VT (verotoxin) production by five samples.

No presente trabalho é descrita a etiologia de um foco de diarréia em leitões de creche em uma granja de ciclo completo. Os exames laboratoriais mostraram o envolvimento simultâneo de E. coli, rotavirus, picobirnavírus e Cryptosporidium parvum. Em-células VERO três amostras de E. coli produziram LT(toxina termolábil) e cinco VT (verotoxina).