RESUMO Objetivou-se, por meio do presente estudo, avaliar o método de colheita farmacológica de sêmen com sondagem uretral, em machos de onças-pardas (Puma concolor) mantidos em cativeiro. A técnica proposta (Cat; N=3) foi comparada com a eletroejaculação (EE; N=4). Para a colheita farmacológica, utilizou-se medetomidina para induzir a liberação de sêmen na uretra e sonda uretral para gatos, sem janela lateral, para colheita do sêmen por capilaridade. O método foi eficaz em todos os animais usados. Por meio dessa técnica, colheram-se amostras com menor volume (106,7±30,5aµL) e maior concentração (524,1±54,3b x 106 espermatozoides/mL) em relação à EE (450,0±0,1bµL e 205,0±141,8a x 106 espermatozoides/mL). As avaliações de vigor, motilidade e patologia espermática demonstraram que a técnica não afeta a qualidade do sêmen em relação à EE (P>0,05). Dessa forma, o método proposto consiste em uma técnica mais prática e eficiente para a colheita de sêmen com boa qualidade, dispensando o eletroejaculador.

ABSTRACT The aim of this study was to evaluate the pharmacological semen collection method with urethral catheterization (CT) in captive cougar (Puma concolor) males. The pharmacological method (CT; N= 3) was compared to the electroejaculation technique (EE; N= 4). For CT collection, medetomidine was administrated to induce semen release using a tomcat catheter inserted into the urethra to collect by capillarity. The proposed method was efficacious on all animals used. Through the CT method, semen collected yielded smaller volume (106,7±30,5aµL) and higher concentration (524,1±54,3b x 106sperm/mL) compared to EE (450,0±0,1bµL and 205,0±141,8a x 106 sperm /mL). Evaluations of vigor, motility and sperm pathology demonstrated that CT does not affect semen quality when compared to EE (P> 0.05). Thus, the proposed method consists of a more practical and efficient technique for semen collection with good quality, eliminating the need for eletroejaculation.

Objetivou-se com o presente estudo monitorar o comportamento do uso territorial de um macho de onça parda, capturado no Parque Estadual da Serra do Brigadeiro (PESB), por meio da telemetria, usando colar de GPS via Satélite Globalstar. O monitoramento de doze meses resultou na transmissão via satélite de 328 localizações, as quais sugerem que a área utilizada por esse felino seja de aproximadamente 610km2, sendo esta a maior área já descrita para a espécie. Foi possível observar que apenas 20% das localizações ocorreram dentro da área do PESB. O comportamento territorial observado neste trabalho mostra a vulnerabilidade do animal a ações antrópicas, como a caça, atropelamentos e o contato com patógenos entre animais silvestres e domésticos. Observou-se também que duas rodovias configuram barreiras artificiais para o deslocamento do animal, possivelmente impedindo o fluxo gênico entre o PESB e dois outros importantes parques. Com os dados levantados é possível a sugestão de ações de manejo, como a criação de uma conectividade efetiva entre o PESB, o Parque Estadual do Rio Doce e o Parque Nacional do Caparaó, o que poderia contribuir para o intercâmbio genético entre as populações e, dessa forma, favorecer a conservação não somente da onça parda.

We aimed to study the territorial behavior of a male cougar, captured in the Parque Estadual da Serra do Brigadeiro (PESB), using a GPS collar and the Globalstar Satellite. Monitoring the animal for nine months we obtained 328 locations. The data suggest that the area used by this feline is approximately 610km2, the largest area described for this specie. It was also possible to observe that only 20% of the locations of the animal occurred within the PESB area. The territorial behavior observed in this study shows the vulnerability of the animal to human activities such as hunting, road kill and contact with pathogens from domestic animals. It was also observed that two roads constitute artificial barriers for animal movement, possibly preventing gene flow between the PESB and two other parks. The creation of an effective connectivity between the PESB, the Parque Estadual do Rio Doce and the Parque Nacional do Caparaó is critical to ensure genetic exchange between the animal populations and thereby promote their conservation.

Tendo em vista a transmissão venérea da leishmaniose visceral do cão para a cadela, o objetivo deste estudo foi verificar se a superfície peniana e o esmegma de cães infectados poderiam ser a fonte de parasitas para a fêmea. Amostras de sêmen e esmegma de 12 cães infectados com Leishmania infantum foram submetidas à identificação do DNA do parasita por PCR. As incidências de positividade no sêmen (41,7%) e no esmegma (50,0%) foram semelhantes (P>0,05; teste exato de Fisher), sendo 58,3% dos cães positivos para sêmen e/ou esmegma. A positividade para sêmen e esmegma juntos ocorreu em 33,3%, mas em 8,3% dos casos apenas no sêmen, e em 16,7% apenas no esmegma, o que revela uma concordância moderada entre os testes (K=0,5; índice Kappa). Conclui-se que a Leishmania infantum está presente no esmegma de cães contaminados, podendo ser a fonte de parasitas para o sêmen e a cadela.

Considering the venereal transmission of visceral leishmaniasis from dogs to bitches, the aim of this study was to verify if the penile surface and smegma from infected dogs can be the source of parasites in bitches. Twelve Leishmania infantum infected dogs had semen and smegma samples collected for submission to PCR identification of the DNA of the parasite. Semen (41.7%) and smegma (50.0%) have similar positive incidence (P>0.05; Fisher's exact test), with 58.3% of the dogs positive for semen and/or smegma samples. The proportion of positivity for both semen and smegma was 33.3%, but 8.3% was positive only for semen, and 16.7% only for smegma, revealing a moderate agreement between tests (K=0.5; Kappa index). It was concluded that Leishmania infantum is present in the smegma of contaminated dogs and it can be a source of parasites for the semen and the bitch.

O objetivo deste trabalho foi adequar a qualidade do leite cru refrigerado de cinco propriedades leiteiras aos parâmetros microbiológicos e de contagem de células somáticas (CCS) estabelecidos pela legislação estadual do Paraná. As propriedades leiteiras estudadas foram monitoradas durante sete meses após a implantação de boas práticas de ordenha e de recomendações quanto ao tratamento de mastites e à manutenção e higienização de equipamentos de ordenha. Antes da adoção das recomendações, a contagem bacteriana total (CBT) média era de 1,36 x 10(6)UFC/mL e a CCS média, de 1,87 x 10(6)/mL. Todas as propriedades atingiram os padrões estabelecidos para CBT, com redução média de 93,4%. A redução de CCS foi, em média, 74,3%. Apenas três propriedades atingiram os valores estabelecidos para CCS. As demais atingiram valores muito próximos ao limite. A adequação aos padrões de CBT e CCS gerou aumento na renda mensal, em razão do pagamento por qualidade do produto, entre R$120,00 e R$828,00, de acordo com a produção de cada propriedade.

The aim of this work was to adequate raw refrigerated milk quality of five dairy farms to microbiological and somatic cell count (SCC) standards established by Paraná state legislation. The dairy farms were monitored during seven months after the implementation of good milking practices and recommendations regarding mastitis treatment, and milking equipment maintenance and hygiene. Before the adoption of recommendations, the average total bacterial count (TBC) was 1.36 x 10(6)CFU/mL and average SCC was 1.87 x 10(6)/mL. All farms reached established standards for TBC, originating an average reduction of 93.4%. Average SCC reduction was 74.3%. Only three farms reached established values for SCC. The remaining reached values near the limits. The suitability of TBC and SCC produced an increase in the monthly income due to payment quality reward of the dairy product, between R$120.00 and R$828.00, according to each farm production.

O presente trabalho consiste no primeiro relato de criptorquidismo em uma jaguatirica, adulta, de vida livre. Para a captura foram empregadas armadilhas com desarme de guilhotina, usando como isca vísceras de bovino. O animal foi contido quimicamente por meio de dardos anestésicos e mantido sob anestesia, utilizando a associação de cloridrato de quetamina e cloridrato de xilazina. Durante o exame andrológico, observou-se que o testículo esquerdo localizava-se subcutâneo, próximo à região inguinal, caracterizando-se criptorquidismo unilateral. Esse testículo apresentava-se flácido, com volume de 2,57mL, enquanto o testículo contralateral apresentava consistência firme e volume de 11,50mL. A área ocupada pelas espículas penianas mostrou-se compatível com a de animais reprodutores. O criptorquidismo é uma condição hereditária ligada à baixa variabilidade genética, já relatada em felinos silvestres consanguíneos. Nesse sentido, devido ao crescente isolamento populacional em fragmentos florestais, este achado torna-se preocupante, uma vez que pode ser indicativo de endogamia em populações de jaguatiricas de vida livre.

This paper is the first report of unilateral cryptorchidism in an adult wildlife ocelot, captured in Parque Estadual do Rio Doce. Cage traps were used to capture the animal, using bovine offal as bait. The animal was anesthetized with anesthetic darts and kept under anesthesia through a combination of ketamine and xylazine. The andrologycal examination showed that the left testicle was located subcutaneously near the inguinal region. In this case of unilateral cryptorchidism, the testis was soft and had a volume of 2.57mL, while the contralateral testis had a firm consistency and volume of 11.50mL. The length of the region occupied by the penile spikes was similar to other breeding animals. Cryptorchidism is an inherited condition linked to low genetic variability previously reported in consanguineous wild cats. Due to the increasing isolation of wild population in forest fragments, this finding is concerning because it can be indicative of inbreeding in wild ocelot populations.

Ourinhos is situated in one of the major sugar cane producing regions in the State of São Paulo, where the manual harvesting is generally preceded by burning the foliage, resulting in large quantities of aerosols being emitted into the atmosphere. A one- month pilot study was executed during August 2010, to characterize the effects of those emissions on the atmosphere, considering the local circulation and the consequences for the region. The plumes were tracked by IPMet´s two S-band Doppler radars, also deploying a large range of meteorological, physical and chemistry instrumentation: a mobile Lidar with Raman channel to observe elevated layers and the type of aerosols, a medium-sized Sodar, as well as 6 Automatic Weather Stations spread through town. Various gases and particulates were also sampled, providing the atmospheric chemistry data base and thus documenting the impact on the region. This paper highlights a case study on 26 August, when a plume was tracked by TITAN Radar Software from the start of the fire, moving southwards at 14-17 km.h-1 with the winds at about 3,5 km above ground level, until it reached Ourinhos 2h15min later, where it was observed by the Raman Lidar and also detected by the aerosol and gas samplers. The high aerosol load of the atmosphere was confirmed by hourly mean values of AOD varying between 0,265 and 0,288 until 07:00 LT, after which they increased to 0,433 by 09:00 LT, as well as hourly mean backscatter profiles. Hourly values of the Lidar Ratio identified the aerosols as biomass burning products, also confirmed through the analysis of gas and aerosol samples simultaneously collected at the Lidar site.

The purpose of the present study was to determine the frequency of hepatitis B virus (HBV) markers in families of HBsAg-positive patients with chronic liver disease. Serum anti-HBc, HBsAg and anti-HBs were determined by enzyme immunoassay and four subpopulations were considered: genetically related (consanguineous) and non-genetically related (non-consanguineous) Asian subjects and genetically related and non-genetically related Western subjects. A total of 165 and 186 relatives of Asian and Western origin were enrolled, respectively. The occurrence of HBsAg and anti-HBs antibodies was significantly higher (P < 0.0001) in family members of Asian origin (81.8%) than in family members of Western origin (36.5%). HBsAg was also more frequent among brothers (79.6 vs 8.5%; P < 0.0001), children (37.9 vs 3.3%; P < 0.0001) and other family members (33.9 vs 16.7%; P < 0.0007) of Asian than Western origin, respectivelly. No difference between groups was found for anti-HBs, which was more frequently observed in fathers, spouses and other non-genetic relatives. HBV infection was significantly higher in children of Asian than Western mothers (P < 0.0004). In both ethnic groups, the mothers contributed more to their children's infection than the fathers (P < 0.0001). Furthermore, HBsAg was more frequent among consanguineous members and anti-HBs among non-consanguineous members. These results suggest the occurrence of vertical transmission of HBV among consanguineous members and probably horizontal sexual transmission among non-consanguineous members of a family cluster. Thus, the high occurrence of dissemination of HBV infection characterizes family members as a high-risk group that calls for immunoprophylaxis. Finally, the study showed a high familial aggregation rate for both ethnic groups, 18/19 (94.7%) and 23/26 (88.5%) of the Asian and Western origin, respectively.

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% 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% of patients with acute hepatitis and in 31% of donors. GBV-C/HGV was detected in 9% of patients with hepatitis, and in 10% 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.

Brazil is a country of continental dimension with a population of different ethnic backgrounds. Thus, a wide variation in the frequencies of hepatitis C virus (HCV) genotypes is expected to occur. To address this point, 1,688 sequential samples from chronic HCV patients were analyzed. HCV-RNA was amplified by the RT-PCR from blood samples collected from 1995 to 2000 at different laboratories located in different cities from all Brazilian States. Samples were collected in tubes containing a gel separator, centrifuged in the site of collection and sent by express mail in a refrigerated container to Laboratório Bioquímico Jardim Paulista, São Paulo, SP, Brazil. HCV- RNA was extracted from serum and submitted to RT and nested PCR using standard procedures. Nested PCR products were submitted to cycle sequencing reactions without prior purification. Sequences were analyzed for genotype determination and the following frequencies were found: 64.9% (1,095) for genotype 1, 4.6% (78) for genotype 2, 30.2% (510) for genotype 3, 0.2% (3) for genotype 4, and 0.1% (2) for genotype 5. The frequencies of HCV genotypes were statistically different among Brazilian regions (P = 0.00017). In all regions, genotype 1 was the most frequent (51.7 to 74.1%), reaching the highest value in the North; genotype 2 was more prevalent in the Center-West region (11.4%), especially in Mato Grosso State (25.8%), while genotype 3 was more common in the South (43.2%). Genotypes 4 and 5 were rarely found and only in the Southeast, in São Paulo State. The present data indicate the need for careful epidemiological surveys throughout Brazil since knowing the frequency and distribution of the genotypes would provide key information for understanding the spread of HCV.

Occult hepatitis B virus (HBV) infection has been reported as cases in which HBV DNA was detected despite the absence of any HBV serological markers or in cases in which anti-HBc antibody was the sole marker. The aim of the present study was to determine, using the polymerase chain reaction (PCR), whether HBV infection occurs in hepatitis C and non-A-E hepatitis patients without serological evidence of hepatitis B infection in São Paulo State. Two different populations were analyzed: 1) non-A-E hepatitis patients, including 12 patients with acute and 50 patients with chronic hepatic disorders without serological evidence of infection with known hepatitis viruses; 2) 43 patients previously diagnosed as hepatitis C with positive results for anti-HCV and HCV RNA. Among hepatitis C patients, anti-HBc was detected in 18.6% of the subjects. Three different sets of primers were employed for HBV DNA detection by nested PCR, covering different HBV genes: C, S and X. HBV-DNA was not detected in any sample, whereas the positive controls did produce signals. The lack of HBV DNA detection with these pairs of primers could be due to a very low viral load or to the presence of mutations in their annealing sites. The latter is unlikely as these primers were screened against an extensive dataset of HBV sequences. The development of more sensitive methods, such as real time PCR, to detect circular covalent closed DNA is necessary in order to evaluate this question since previous studies have shown that cryptic hepatitis B might occur.

Avaliou-se o potencial adjuvante do cloreto de dimetildioctadecilamônio (DDA cloreto) em estimular as respostas imune humoral e celular, do tipo hipersensibilidade cutânea tardia (DTH), em cobaios que receberam preparações de antígeno contendo o herpesvírus bovino tipo 1 (BHV-1) inativado. Os animais foram vacinados com o BHV-1 em cinco diferentes formulações, representadas pelos grupos: A- adjuvante completo / incompleto de Freund, B- hidróxido de alumínio (Al(OH)3), C- DDA cloreto, D- associação Al(OH)3 / DDA cloreto e E- BHV-1 em solução aquosa sem adjuvante. Os animais do grupo F, grupo-controle negativo, foram inoculados apenas com meio de cultivo celular. Os maiores títulos de anticorpos neutralizantes do BHV-1, expressos em log2, foram obtidos nos grupos D, A e C, com títulos de 6,25, 6,0 e 5,25, respectivamente. Na avaliação da DTH, os maiores aumentos na espessura da dobra da pele foram observados nos grupos A (2,4mm), C (1,8mm) e D (1,1mm). O DDA cloreto, utilizado tanto de forma isolada quanto em associação, determinou a potencialização das respostas imunológicas humoral e celular de cobaios imunizados com o BHV-1 inativado.

The adjuvanticity of dimethyl dioctadecyl ammonium chloride (DDA chloride) to induce humoral and cell mediated (delayed type hypersensitivity - DTH -) immune responses was assessed in guinea pigs that received antigen preparations containing inactivated bovine herpesvirus type 1 (BHV-1). The animals were vaccinated with five different formulations, containing BHV-1 represented by groups: A- complete/incomplete Freund adjuvant, B- aluminum hydroxide (Al(OH)3), C- DDA chloride, D- association Al(OH)3/DDA and E- BHV-1 in aqueous solution without adjuvant. Group F was the negative control group and the animals received only cell culture medium. The higher titers of BHV-1 neutralizing antibodies, expressed in log2, were obtained in groups D, A and C with values of 6.25, 6.0 and 5.25, respectively. In the DTH assessment, the higher increase in skin thickness was observed in group A (2.4mm), C (1.8mm) and D (1.1mm). These results showed that DDA chloride used alone or in association to other substance boosted the humoral and cell mediated immune responses in guinea pigs immunized with inactivated BHV-1.

Parvovirus B19 has been associated by some investigators with cases of severe hepatitis. The aim of the present study was to determine the presence of active parvovirus B19 infection among 129 Brazilian patients with non-A-E hepatitis. The patients were assayed for antibodies against parvovirus B19, IgM class, by ELISA. In IgM-positive cases, parvovirus B19 DNA was assayed by PCR in serum and liver tissue and parvovirus VP1 antigen in liver tissue was assayed by immunohistochemistry. Antibodies against parvovirus B19, IgM class, were detected in 3 (2.3%) of 129 patients with non-A-E hepatitis. Previous surgery and blood transfusions were reported by these 3 patients. One patient was a 56-year-old female with severe hepatitis, with antimitochondrial antibody seropositivity and submassive necrosis at liver biopsy, who responded to corticosteroid therapy. Strong evidence for active parvovirus B19 infection was found in this patient, with parvovirus B19 DNA being detected by PCR in liver tissue. Furthermore, parvovirus VP1 antigen was also detected in liver tissue by immunohistochemistry. The other two IgM-positive patients were chronic hepatitis cases, but active infection was not proven, since neither viral DNA nor antigen were detected in their liver tissues. This and other reports suggest a possible relation between parvovirus B19 infection and some cases of hepatitis.

O vírus da Hepatite G ou vírus GB-C é um novo vírus recentemente descoberto em casos de hepatites não A-E. Neste estudo, casos de hepatite crônica não-B, não-C foram testados com uma reação de amplificação específica para GBV-C. Este vírus foi detectado em 1 entre os 13 casos estudados. Este paciente apresentava insuficiência hepática severa, tinha habitado por vários anos na Amazônia Ocidental e nenhuma outra causa para este quadro clínico havia sido relatada. O impacto da descoberta deste novo agente está ainda sendo investigado. O estudo da prevalência deste vírus entre pacientes com hepatite crônica e entre indivíduos sadios (como, por exemplo, doadores de sangue) fornecerá subsídios para o estabelecimento de sua real patogenicidade.

Hepatitis G virus/ GB virus C is a novel flavivirus recently detected in hepatitis non A-E cases. In this study, the presence of this virus in chronic non-B, non-C hepatitis patients was evaluated using GBV-C specific PCR and this virus was detected in one out of thirteen patients. This patient has presented a severe liver failure, has lived for a long time in the Western Amazon basin and no other cause for this clinical picture was reported. The impact of the discovery of this new agent is still under evaluation throughout the world. The study of the prevalence of this virus among chronic hepatitis patients and healthy individuals (as blood donors) will furnish subside to evaluate its real pathogenicity.

A resposta ao tratamento com interferon em pacientes com hepatite crônica NANB/C tem sido classificada como completa, parcial ou ausente, de acordo com o comportamento da alanino aminotransferase sérica (ALT). Entretanto, uma observação mais detalhada da atividade enzimática tem mostrado que os padrões podem ser mais complexos. O objetivo deste estudo foi descrever o seguimento tardio e os padrões de resposta da ALT em pacientes com hepatite crônica NANB/C tratados com interferon alfa recombinante. Classificamos os tipos de resposta da ALT em 6 padrões e as freqüências observadas foram:I. Resposta completa e persistente = 9 (20,5%); II. Resposta completa com recaída persistente após o IFN = 7 (15,9%); III. Resposta completa com recaída temporária = 5 (11,9%); IV. Normalização temporária e recaída durante o tratamento com IFN = 4 (9,1%); V. Resposta parcial (queda dos níveis iniciais da ALT maior que 50%) = 7 (15,9%); VI. Não resposta = 12 (27,3%). Em conclusão, os padrões de ALT variam durante e após o tratamento e podem ser classificados em pelo menos 6 tipos.

The response to interferon treatment in chronic hepatitis NANB/C has usually been classified as complete, partial or absent, according to the behavior of serum alanine aminotransferase (ALT). However, a more detailed observation of the enzymatic activity has shown that the patterns may be more complex. The aim of this study was to describe the long term follow-up and patterns of ALT response in patients with chronic hepatitis NANB/C treated with recombinant interferon-alpha. A follow-up of 6 months or more after interferon-a was achieved in 44 patients. We have classified the serum ALT responses into six patterns and the observed frequencies were as follows: I. Long term response = 9 (20.5%); II. Normalization followed by persistent relapse after IFN = 7 (15.9%); III. Normalization with transient relapse = 5 (11.9%); IV. Temporary normalization and relapse during IFN = 4 (9.1%); V. Partial response (more than 50% of ALT decrease) = 7 (15.9%); VI. No response = 12 (27.3%). In conclusion, ALT patterns vary widely during and after IFN treatment and can be classified in at least 6 types.