Microbial source tracking and antimicrobial resistance in one river system of a rural community in Bahia, Brazil / Rastreamento de fontes microbianas e resistência antimicrobiana em um sistema fluvial de uma comunidade rural da Bahia, Brasil

Use of antibiotics inevitably leads to antimicrobial resistance. Selection for resistance occurs primarily within the gut of humans and animals as well as in the environment through natural resistance and residual antibiotics in streams and soil. We evaluated antimicrobial resistance in Gram negative bacteria from a river system in a rural community in Bahia, Brazil. Water was collected from the Jiquiriçá and Brejões rivers and the piped water supply. Additionally, stools were collected from a random sample of residents, cows, pigs and horses near the river. The samples were screened for bacteria resistant to ciprofloxacin, cefotaxime, and meropenem and identified biochemically at the genus and species levels. Microbial source tracking demonstrated that ruminant and human fecal contamination increased as the rivers neared the village center and decreased after the last residence. Antibiotic bacteria were identified from all samples (n = 32). No bacteria were resistant to carbapenems, but the majority of the enterobacteria were resistant to ciprofloxacin, even though this class of antibiotics is not commonly used in food animals in this region. Considering these facts, together with the pattern of human fecal contamination, a human source was considered most likely for these resistant isolates.

O uso de antibióticos inevitavelmente leva à resistência antimicrobiana. A seleção para resistência antimicrobiana ocorre principalmente no intestino de seres humanos e animais, bem como no meio ambiente, através da resistência natural e resíduos de antibióticos nos esgotos e no solo. Avaliamos a resistência antimicrobiana em bactérias Gram-negativas de um sistema fluvial em uma comunidade rural da Bahia, Brasil. A água foi coletada nos rios Jiquiriçá e Brejões e no abastecimento de água encanada. Além disso, foram coletadas amostras randomizadas de fezes de moradores, vacas, porcos e cavalos próximos ao rio. As amostras foram triadas para bactérias resistentes à ciprofloxacina, cefotaxima e meropenem e identificadas bioquimicamente nos níveis de gênero e espécie. O rastreamento de fontes microbianas demonstrou que a contaminação fecal de ruminantes e humanos aumentou à medida que os rios se aproximavam do centro da vila e diminuía após a última residência. Bactérias resistentes a antibióticos foram identificadas em todas as amostras (n = 32). Nenhuma bactéria demonstrou ser resistente aos carbapenêmicos testados, contudo, foi encontrado enterobactérias resistentes à ciprofloxacina, ainda que essa classe de antibióticos não seja comumente usada na medicina veterinária dos animais dessa região. Considerando esses fatos, juntamente com o padrão de contaminação fecal avaliado, a fonte de contaminação humana foi considerada a mais provável na interação desses isolados resistentes.

Microbial source tracking and antimicrobial resistance in one river system of a rural community in Bahia, Brazil

Abstract Use of antibiotics inevitably leads to antimicrobial resistance. Selection for resistance occurs primarily within the gut of humans and animals as well as in the environment through natural resistance and residual antibiotics in streams and soil. We evaluated antimicrobial resistance in Gram negative bacteria from a river system in a rural community in Bahia, Brazil. Water was collected from the Jiquiriçá and Brejões rivers and the piped water supply. Additionally, stools were collected from a random sample of residents, cows, pigs and horses near the river. The samples were screened for bacteria resistant to ciprofloxacin, cefotaxime, and meropenem and identified biochemically at the genus and species levels. Microbial source tracking demonstrated that ruminant and human fecal contamination increased as the rivers neared the village center and decreased after the last residence. Antibiotic bacteria were identified from all samples (n = 32). No bacteria were resistant to carbapenems, but the majority of the enterobacteria were resistant to ciprofloxacin, even though this class of antibiotics is not commonly used in food animals in this region. Considering these facts, together with the pattern of human fecal contamination, a human source was considered most likely for these resistant isolates.

Resumo O uso de antibióticos inevitavelmente leva à resistência antimicrobiana. A seleção para resistência antimicrobiana ocorre principalmente no intestino de seres humanos e animais, bem como no meio ambiente, através da resistência natural e resíduos de antibióticos nos esgotos e no solo. Avaliamos a resistência antimicrobiana em bactérias Gram-negativas de um sistema fluvial em uma comunidade rural da Bahia, Brasil. A água foi coletada nos rios Jiquiriçá e Brejões e no abastecimento de água encanada. Além disso, foram coletadas amostras randomizadas de fezes de moradores, vacas, porcos e cavalos próximos ao rio. As amostras foram triadas para bactérias resistentes à ciprofloxacina, cefotaxima e meropenem e identificadas bioquimicamente nos níveis de gênero e espécie. O rastreamento de fontes microbianas demonstrou que a contaminação fecal de ruminantes e humanos aumentou à medida que os rios se aproximavam do centro da vila e diminuía após a última residência. Bactérias resistentes a antibióticos foram identificadas em todas as amostras (n = 32). Nenhuma bactéria demonstrou ser resistente aos carbapenêmicos testados, contudo, foi encontrado enterobactérias resistentes à ciprofloxacina, ainda que essa classe de antibióticos não seja comumente usada na medicina veterinária dos animais dessa região. Considerando esses fatos, juntamente com o padrão de contaminação fecal avaliado, a fonte de contaminação humana foi considerada a mais provável na interação desses isolados resistentes.

Assessment of surface temperatures of buffalo bulls (Bubalus bubalis) raised under tropical conditions using infrared thermography / Avaliação de temperaturas superficiais de touros bubalinos (Bubalus bubalis) criados em ambiente tropical com uso da termografia infravermelha

This paper aimed to evaluate the surface temperatures of buffalo bulls using infrared thermography, considering four distinct anatomical parts over time, and to correlate surface temperatures and thermal comfort indexes. The humid tropical climate (Kõppen's Afi) was predominant in the research station where the experiment was performed and the trial lasted from April to August. Ten bulls (n=10) were evaluated every 25 days (morning: 6:00-9:00; afternoon: 12:00-15:00) and the parameters assessed were respiratory rate (RR), rectal temperature (RT), and the thermograms of surface temperature for orbital area (ORB), right flank (RF), left flank (LF) and scrotum (SCR). Climatological data was continuously monitored and the Temperature and Humidity Index (THI) and the Index of Comfort of Benezra (ICB) were calculated. The average values of THI were ≥78, and significant differences between shifts were observed (P<0.05). The ICB ranged from 1.96 to 2.25 and significant differences were observed for shifts and throughout the months (P<0.05). The averages of surface temperatures were RT=38.2±0.5°C, ORB=36.1±0.8°C, LF=33.5±2.5°C, RF=35.4±1.7ºC and SCR=33.3±1.1°C, which exhibited significant differences for shifts and throughout the months (P<0.05). Positive correlations were obtained between THI and ORB (0.72), RF (0.77), LF (0.75) and SCR (0.41) (P<0.0001). The maximum temperature of ORB showed the highest correlation with RT (0.58, P<0.0001). Therefore, the surface temperatures are subject to climatic variations and increase throughout the day, due to the variation in thermal comfort indexes, and the maximum ORB temperature was the parameter most related to rectal temperature. Lastly, the results indicate that IRT may be a useful non-invasive and accurate tool to detect the variations in ORB, LF, RF and SCR temperature in buffalo bulls.

O presente trabalho visou avaliar as temperaturas superficiais de diferentes regiões anatômicas de búfalos ao longo do tempo, por meio da termografia infravermelha, e correlacioná-las a índices bioclimatológicos de conforto térmico. O ensaio foi realizado em região de clima tropical úmido (Afi de Kõppen), de abril a agosto. Dez touros (n=10) foram avaliados a cada 25 dias (manhã: seis-nove horas; tarde: 12-15h), quanto à frequência respiratória (FR), temperatura retal (TR) e imagens termográficas da órbita ocular (ORB), flanco direito (FLd), flanco esquerdo (FLe) e escroto (ESC). Os dados climatológicos foram ininterruptamente monitorados, e calculados o índice de temperatura e umidade (ITU) e o índice de conforto de Benezra (ICB). O ITU foi ≥78, com diferença entre turnos (P<0,05). Já o ICB variou de 1,96 a 2,25 e apresentou diferenças ao longo dos meses e entre turnos (P<0,05). As temperaturas observadas foram de TR=38,2±0,5ºC, ORB=36,1±0,8ºC, FLd=33,5±2,5ºC, FLe=35,4±1,7ºC e ESC=33,3±1,1ºC, as quais variaram significativamente ao longo dos meses e entre turnos (P<0,05). O ITU apresentou correlações positivas com ORB (0,72), FLd (0,77), FLe (0,75) e ESC (0,41) (P<0,0001). A temperatura máxima de ORB apresentou a maior correlação com a TR (0,58; P<0,0001). Portanto, as temperaturas superficiais dos animais sofrem interferências das variações climáticas e se elevam ao longo do dia, devido à variação nos índices de conforto térmico; a temperatura máxima de ORB foi o parâmetro mais condicionado à temperatura retal. Também, as oscilações de temperatura de superfície de ORB, FLd, FLe e ESC podem ser aferidas em bubalinos com o uso da termografia infravermelha, de modo preciso e não invasivo.

Transporte de oócitos bovinos em meio de maturação por diferentes períodos de tempo sem controle da atmosfera gasosa / Transport of bovine oocytes in maturation medium for different periods of time whithout controlled gaseous atmosphere

Avaliou-se a viabilidade do transporte de oócitos em meio quimicamente definido, e analisou-se a necessidade da adição ou não de hormônios neste meio. Os oócitos do grupo-controle (0h) foram maturados por 24h em estufa de CO2, e os dos grupos experimentais foram transportados em incubadora portátil. No experimento I, as taxas de clivagem foram similares (P>0,05) para os grupos 0h (59,7 por cento), 3h (53,5 por cento) e 9h (48,8 por cento), e houve redução nos grupos 6h (46,1 por cento) e 12h (43,8 por cento). Essas taxas foram semelhantes entre os grupos 3h, 6h, 9h e 12h. A produção de blastocistos não foi diferente (P>0,05) para os grupos 0h (38,0 por cento), 3h (32,3 por cento), 6h (27,3 por cento) e 9h (24,8 por cento), e houve redução no grupo 12h (18,9 por cento). Essas taxas foram semelhantes entre os grupos 6h, 9h e 12h. No experimento II, não houve diferença (P>0,05) entre as taxas de clivagem para os grupos 0h (71,4 por cento), 3h (70,3 por cento), 6h (56,0 por cento) com hormônios, e os grupos 3h (64,8 por cento) e 6h (54,1 por cento) sem hormônios. A produção de blastocistos foi similar (P>0,05) para os grupos 0h (46,1 por cento), 3h com hormônios (45,8 por cento) e 3h sem hormônios (41,1 por cento), porém houve redução nos grupos 6h com hormônios (35,5 por cento) e 6h sem hormônios (33,5 por cento). Essas taxas foram semelhantes entre os grupos 3h sem hormônios e 6h com e sem hormônios. Estes resultados indicam que é possível otransporte de oócitos bovinos por um período de até nove horas, e que a adição de hormônios neste meio não influencia os índices de clivagem e de blastocistos.

The viability of the transport of the bovine oocytes was evaluated in chemically defined medium and the need for the addition or not of hormones in this medium was analyzed. The oocytes in the control group (0h) were matured for 24h in CO2 incubator, and in experimental groups they were transported in portable incubator. In experiment I, the cleavage rates were similar (P>0.05) to the groups 0h (59.7 percent), 3h (53.5 percent), and 9h (48.8 percent), but they decreased in groups 6h (46.1 percent) and 12h (43.8 percent), however, these rates were similar among the groups 3h, 6h, 9h, and 12h. The production of blastocysts was not different (P>0.05) for groups 0h (38.0 percent), 3h (32.3 percent), 6h (27.3 percent), and 9h (24.8 percent), but there was a reduction in the 12h group (18.9 percent). These rates were similar among the groups 6h, 9h and 12h. In experiment II, no significant difference (P>0.05) was observed among the rates of cleavage for the groups 0h (71.4 percent), 3h with (70.3 percent) and without hormones (64.8 percent), and 6h with (56.0 percent) and without hormones (54.1 percent). The production of blastocysts was similar (P>0.05) for groups 0h (46.1 percent) and 3h with (45.8 percent) and without hormones (41.1 percent), but decreased in groups 6h with (35.5 percent) and without hormones (33.5 percent). These rates were similar among the groups 3h without, 6h with and without hormones. These results indicate the possibility of the transport of bovine oocytes up to 9h, and the addition of hormones in this medium does not influence the rates of cleavage and blastocysts.

Efeito do estradiol e da progesterona no desenvolvimento e na qualidade de embriões bovinos produzidos in vitro / Effect of estradiol and progesterone on development and quality of bovine embryos produced in vitro

Avaliaram-se o desenvolvimento e a qualidade de embriões bovinos, cocultivados com células epiteliais do oviduto bovino (CEOBs) expostas ou não ao estradiol e à progesterona. Os ovócitos foram maturados in vitro por 24h e, então, fertilizados utilizando-se sêmen congelado, em estufa de CO2 a 5 por cento e 38,5ºC. As CEOBs foram cultivadas em TCM-199 com ou sem estradiol (E2) (24 horas), nas mesmas condições da maturação e fertilização in vitro (MIV e FIV), e, em seguida, adicionadas aos diferentes grupos em CR2 com ou sem progesterona (P4) (G1=P4+E2); (G2=E2); (G3=P4) e (G4=controle). Após 18h da FIV, as células foram cultivadas nos diferentes sistemas. Nenhuma diferença (P>0,05) foi observada nas taxas de clivagem entre G1, G2 e G4 (53,5 por cento; 56,3 por cento; 51,7 por cento) e nos padrões de blastocistos (BLs) (29,3 por cento; 31,2 por cento, 28,7 por cento). Índices menores (P<0,05) foram obtidos no G3 para ambas as variáveis (34,5 por cento; 16,4 por cento). G1 e G2 apresentaram taxas de eclosão maiores (P<0,05) que os outros grupos (23,3 por cento; 23,2 por cento), sendo G4 (19,3 por cento) diferente de G3 (16,1 por cento). Em G1, G2 e G3, o número de células nos BLs aumentou 125,9; 128,4 e 123,6, respectivamente (P<0,05), em relação ao G4 (112,5). Conclui-se que o tratamento das CEOBs com o E2, nas primeiras 24 horas de cultivo, pode ser usado isoladamente ou em combinação com a progesterona, a fim de melhorar a qualidade de embriões bovinos produzidos in vitro.

The development and quality of bovine embryos co-cultivated with bovine oviduct epithelial cells (BOEC) supplemented or not with estradiol and progesterone were evaluated. Oocytes were matured and fertilized in vitro (MIV/FIV) (5 percentCO2/38.5ºC). The BOEC were cultivated in TCM-199 with or without estradiol (E2) (24h), in the same conditions of MIV/FIV. Presumptive zygotes were transferred to BOEC in suspension after in vitro maturation and fertilization of bovine oocytes with thawed percoll-selected sperm. The zygotes were cultivated in CR2 medium containing or not progesterone (P4) (G1=P4+E2), (G2=E2), (G3=P4), and (G4=control). No significant differences (P>0.05) were found in the cleavage rates among G1, G2, and G4 (53.5 percent, 56.3 percent, and 51.7 percent) as well as in relation to the blastocystis (BL) rates (29.3 percent, 31.2 percent, and 28.7 percent). However, significant differences (P<0.05) were observed in the G3 for both variables (34.5 percent and 16.4 percent). G1 and G2 showed hatching rates higher (P<0.05) than the other groups (23.3 percent; 23.2 percent), being G4 (19.3 percent) different from G3 (16.1 percent) (P<0.01). In the groups G1, G2, and G3, the total cell number of the BL increased (125.9, 128.4, and 123.6) (P<0.05) compared to G4 (112.5). These results demonstrate that the treatment of the BOEC with estradiol in first the 24 hours of culture can be used separately or in combination with the progesterone to improve the quality of bovine embryos produced in vitro.

Prevalence of hepatitis C virus (HCV) infection and HCV genotypes of hemodialysis patients in Salvador, Northeastern Brazil

Hepatitis C virus (HCV) infection has been identified as the major cause of chronic liver disease among patients on chronic hemodialysis (HD), despite the important reduction in risks obtained by testing candidate blood donors for anti-HCV antibodies and the use of recombinant erythropoietin to treat anemia. A cross-sectional study was performed to estimate the prevalence of HCV infection and genotypes among HD patients in Salvador, Northeastern Brazil. Anti-HCV seroprevalence was determined by ELISA in 1243 HD patients from all ten different dialysis centers of the city. HCV infection was confirmed by RT-PCR and genotyping was performed by restriction fragment length polymorphism. Anti-HCV seroprevalence among HD patients was 10.5 percent (95 percent CI: 8.8-12.3) (Murex anti-HCV, Abbott Murex, Chicago, IL, USA). Blood samples for qualitative HCV detection and genotyping were collected from 125/130 seropositive HD patients (96.2 percent). HCV-RNA was detected in 92/125 (73.6 percent) of the anti-HCV-positive patients. HCV genotype 1 (77.9 percent) was the most prevalent, followed by genotype 3 (10.5 percent) and genotype 2 (4.6 percent). Mixed infections of genotypes 1 and 3 were found in 7.0 percent of the total number of patients. The present results indicate a significant decrease in anti-HCV prevalence from 23.8 percent detected in a study carried out in 1994 to 10.5 percent in the present study. The HCV genotype distribution was closely similar to that observed in other hemodialysis populations in Brazil, in local candidate blood donors and in other groups at risk of transfusion-transmitted infection.

HEV, TTV and GBV-C/HGV markers in patients with acute viral hepatitis

The aim of the present study was to evaluate the prevalence of HEV, TTV and GBV-C/GBV-C/HGV in patients with acute viral hepatitis A, B and non-A-C. We evaluated sera of 94 patients from a sentinel program who had acute hepatitis A (N = 40), B (N = 42) and non-A-C (N = 12); 71 blood donors served as controls. IgM and anti-HEV IgG antibodies were detected by enzyme immunoassay using commercial kits. TTV and GBV-C/HGV were detected by nested PCR; genotyping was done by sequencing and phylogenetic analysis. Anti-HEV IgG was present in 38, 10 and 17 percent of patients with hepatitis A, B and non-A-C. Four patients with hepatitis A and 1 with non-A-C hepatitis also had anti-HEV IgM detected in serum. TTV was detected in 21 percent of patients with acute hepatitis and in 31 percent of donors. GBV-C/HGV was detected in 9 percent of patients with hepatitis, and in 10 percent of donors. We found TTV isolates of genotypes 1, 2, 3, and 4 and GBV-C/HGV isolates of genotypes 1 and 2. Mean aminotransferase levels were lower in patients who were TTV or GBV-C/HGV positive. In conclusion, the detection of anti-HEV IgM in some acute hepatitis A cases suggests co-infection with HEV and hepatitis E could be the etiology of a few cases of sporadic non-A-C hepatitis in Salvador, Brazil. TTV genotype 1, 2, 3 and 4 isolates and GBV-C/HGV genotype 1 and 2 strains are frequent in the studied population. TTV and GBV-C/HGV infection does not appear to have a role in the etiology of acute hepatitis.

Comparative study of Hepatitis C virus genotypes 1 and 3 in Salvador, Bahia

Hepatitis C virus displays a high degree of genetic mutation, with considerable heterogeneity, motivating clinical and biomolecular investigations. It is necessary to understand the effects of genotypes on the course of the disease, as well as their peculiarities at the regional level. OBJECTIVE: The study objective was to compare epidemiological, biochemical and histological aspects of hepatitis C virus genotypes 1 and 3 in Salvador, Bahia. STUDY DESIGN: Data were collected retrospectively from outpatient medical records. MATERIALS AND METHODS: 127 patients with positive anti-HCV results were selected, based on detectable RNA-HCV (RT-PCR) of genotypes 1a, 1b and 3a. RESULTS: Thirty-nine (30.7 percent) individuals were infected by subtype 1a, 45 (35.4 percent) by subtype 1b and 43 (33.9 percent) by subtype 3a. Most (73.2 percent) patients were male, with an average age of 47.8 years. The subtype 1b-infected patients had the highest average age (512 ±11.17; P=0.09). The use of illicit injected drugs was more frequent among subtype 3a infected individuals when compared with genotype 1 (6/43; 14 percent and 3/84; 3.6 percent, respectively; P=0,06). No significant differences were found for other epidemiological characteristics. Average values for GT, AST, ALT and ferritin did not differ between the groups (64, 78, 109, 276, respectively). Thyroid dysfunction occurred in 7/30 (23.3 percent) of those infected by genotype 3 (P=0.05). Cryoglobulinemia was also more frequent in this group (5/13, 38 percent, P=0.02). Most patients presented limited necro-inflammatory activity, stages 2 and 3 by the METAVIR Classification. In some cases, dissociation was noticed between inflammatory activity and fibrosis. No significant differences were found in the histopathological findings of the various genotypes. Younger patients had a significantly smaller degree of necrosis in stomatocytosis (P=0.032) and fibrosis (P=0.012). Intense parenchymatous activity and lymphoid follicles were more frequent among alcohol consumers (P=0.06 and P=0.04, respectively). CONCLUSIONS: In Bahia, genotype 3 dissemination seems to be associated with illicit drug use. The disease evolution depends on a function of complex interactions between virus and host. Age and alcohol consumption stand out as important variables in the development of cirrhosis.