Background Cryptococcus gattii-induced cryptococcosis is an emerging infectious disease of humans and animals with worldwide distribution and public health importance due to its significant morbidity and mortality rate. The present study aimed to report a case of pulmonary infection by C. gattii molecular type VGII in State of São Paulo, Brazil.Case presentationA 5-year-old goat showing intermittent dry cough, ruminal tympany, anorexia, fever, tachycardia and tachypnea was presented for necropsy at the Veterinary Hospital of the School of Veterinary Medicine and Animal Sciences, São Paulo University, São Paulo, Brazil. Postmortem examination revealed numerous 2.0–6.0 cm diameter yellow gelatinous pulmonary masses. Tissues were evaluated by a combination of pathological, mycological, and molecular diagnostic techniques. Microscopically, pneumonia granulomatous, multifocal to coalescing, moderate, with many intralesional carminophilic yeasts was observed. The immunohistochemistry and mycological culture confirmed Cryptococcus spp. Internal transcribed spacers and orotidine monophosphate pyrophosphorylase nucleotide differentiation demonstrated that the isolate corresponds to the C. gattii VGII molecular subtype.ConclusionsTo our knowledge, this is the first report of a pulmonary infection in a goat linked to C. gattii molecular type VGII in Southeastern Brazil. Our findings emphasize the need for an active surveillance program for human and animal new infections to improve the current public health policies due to expansion of the epidemiological niche of this important microorganism.

The detection of Bovine Viral Diarrhea virus (BVDV) infection, especially among persistently infected calves (PI), should be performed earlier in order to eliminate the source of the infection and to prevent the spread of the disease in the herd. However, colostrum intake can influence the results of some of the tests used to diagnose the BVDV infection. Therefore, this study evaluated the efficacy of serum neutralization (SN) test in conjunction with reverse transcription-polymerase chain reaction (RT-PCR) for the diagnosis of BVDV infection before colostrum intake. The deliveries of the animals were assisted to select 52 newborn Holstein calves for inclusion in the study. Initially, the whole blood and serum samples were collected from the calves before (T0) and after (T1) the colostrum intake. The calves that were RT-PCR positive at any of the time-points were retested on the 30 th day post birth (T2). The presence of specific antibodies for BVDV was evaluated by SN, and that of viral RNA by the RT-PCR. The BVDV-specific antibodies were observed in the serum of 13.46% (7/52) of the calves at T0 because of fetal infection. At T1, seroconversion was observed in 100% (52/52) of the calves. The geometric mean titers (GMT) of the antibodies for BVDV increased significantly from T0 (14.52) to T1 (2490) (P = 0.0001). Of the four calves that were RT-PCR positive before colostrum intake (T0), two were seronegative and two, seropositive. Of the forty-eight RT-PCR negative calves, five were seropositive. After 30 days post birth, all of the animals tested negative by RT-PCR, thus excluding the possibility of persistent infection. The association observed between the results of the SN and RT-PCR assays at T0 (P = 0.025) could not be observed at T1 (P > 0.05). The SN test before the colostrum intake allowed the detection of fetal infection in the herd; however, this test was ineffective as a diagnostic method after the transfer of passive immunity. The confirmation of the results of the SN assay by those of the RT-PCR was essential for the identification of the infected calves before colostrum intake. Key words: Bovine viral diarrhea. Passive immunity. Persistent infection. Polymerase chain reaction. Serum neutralization. ResumoInfecção causada pelo BVDV, especialmente bezerras persistentemente infectadas (PI), deve ser detectada precocemente para eliminação da fonte de infecção e disseminação da doença no rebanho. No entanto, a mamada de colostro interfere em alguns testes adotados para o diagnóstico da Diarréia Viral Bovina. Assim, esta pesquisa avaliou o uso da soroneutralização (SN) em associação com a reação em cadeia de polimerase precedida da transcrição reversa (RT-PCR) para diagnóstico da infecção pelo BVDV antes da mamada de colostro. Partos foram acompanhados para seleção de 52 bezerras da raça Holandesa. Inicialmente foram coletadas amostras de sangue total e soro de todos os animais antes (T0) e após a mamada do colostro (T1). Os animais positivos no RT-PCR em qualquer momento foram retestados aos 30 ...

Background: Infections are caused by Bovine Viral Diarrhea Virus and still continue to be a worldwide plague in cattle industry. It is responsible for sudden death syndromes in adult cattle with high mortality rates, abortions, acute gastrointestinal and respiratory diseases. The BVDV infection occurs in early pregnancy (40-142 days), in immunosuppressed females or cows results in 100% of persistently infected (PI) calves that are seronegative and asymptomatic at birth. Evidences suggests that BVDV contributes to BRD complex potentiating secondary infections caused by Mannheimia haemolytica e Pasteurella multocida due to its immunosuppressive action. However, the farmers have often associated the respiratory syndrome with other infectious agents. This paper reports the attendance of dairy calves manifesting clinical signs of bronchopneumonia, which led to the screening of the persistently infected animals to control of the BVDV infection in the herd.Materials, Methods & Results: During the technical assistance, ten calves manifesting bronchopneumonia were selected to trans-tracheal lavage (TL) in order to identify possible infectious agents. Reverse transcription polymerase chain reaction (RT-PCR) detected the presence of BVDV in two heifers. Pasteurella multocida was the unique bacterial agent isolated from TL (5/10, 50%). These data motivated the technical team and producers to investigate the PI screening by direct enzyme-linked immunosorbent assay from biopsies of the ear edge. The screening of PI’s detected 29 positive within of 2,342 animals tested (1.23%). The re-test of positive was performed only in 24 animals due to the cull of five bovine with severe bronchopneumonia and diarrhea, confirming 18 persistently infected calves (18/24; 75%). Finally, in all PI’s live dams were tested. It was observed four positive adult animals. One grand dam was live and tested, but it had negative result for direct enzyme-linked immunosorbent test. The rate of PI’s considering the whole herd was 0.81% (22/3,700 animals).Discussion: The involvement of BVDV in the etiology of bronchopneumonia was confirmed by detection of the virus in trans-tracheal lavages in two calves by RT-PCR. The susceptibility for Pasteurella multocida infection could be promoted by BVDV prime-infection, considering that immunossupressive nature of BVDV is a critical factor in the interaction with others viruses and bacteria. At this time, we are aware about any report about the detection of BVDV in trans-tracheal lavages. These findings culminated with the screening of PI animals in the herd, detecting rates of 0.81%. The intensive vaccination and colostrum management of this farm could protected the herd against BVDV, however others facts facilitated the introduction of the virus in the herd. This research was conduced in a high-production dairy farm with around 3,700 animals raised in an open herd, in which some of cows with high genetic potential were transferred for embryo collection in the state of Paraná, Brazil; resulting in the addition of the calves to the herd by others routes. Moreover, the farm used for many years vaccine containing only BVDV-1, which may have favored the entry and spread of BVDV-2 or BVDV-3 in the herd. This research showed the presence of BVDV in trans-tracheal lavage of heifers with bronchopneumonia by RT-PCR. This fact points to the need of BRD control programs that include detection of PI animals.

Um surto de fotossensibilização hepatógena por ingestão de Brachiaria decumbens é relatado em bovinos da raça Nelore. Após um lote de vacas e novilhas (n=320) ser transferido para um pasto de Brachiaria decumbens, vários animais começaram a apresentar diarréia, 15 apresentaram lesões de pele, que estava ressequida, enrugada com fissuras e grande quantidade de crostas, e 80 animais morreram. Foram coletadas amostras de sangue, urina e fezes de alguns animais. O hemograma apresentou alterações inespecíficas e o exame de urina e fezes estava dentro dos parâmetros aceitáveis para a espécie. Foi realizada a necropsia de um animal. O fígado apresentava coloração amarelo esverdeada. Microscopicamente havia pericolangite linfocitária multifocal leve, com tumefação difusa de hepatócitos e hepatócitos com vacuolização acentuada, bilestase difusa, presença de cristais birrefringentes e macrófagos espumosos. No rim observou-se nefrose tubular multifocal leve a moderada com presença de gotículas hialinas na luz dos túbulos, e no lúmen havia células inflamatórias, principalmente macrófagos. Após o sombreamento de algumas áreas do pasto e o inicio do tratamento com hepatoprotetor, antibióticos e limpeza das feridas os animais apresentaram melhora e não ocorreram outros casos de fotossensibilização.O diagnóstico de fotossensibilização hepatógena foi baseado nos sinais clínicos, nas lesões histológicas hepáticas e renais e no relato da transferência para um pasto de Brachiaria decumbens.

After vaccination, vaccine components must activate the immune response, but the ideal vaccine should not result in undesirable effects in cattle. The aim of this study was to evaluate the inflammatory and humoral responses and adverse reactions induced by three adjuvanted commercial vaccines against bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BHV-1). Holstein heifers (n = 35) were divided into four groups by adjuvant compounds: Vaccine A (Alum; n=9), Vaccine B (Oil-in-water; n=10), Vaccine C (Amphigen/Quil A cholesterol and dimethyl-dioctadecyl ammonium (DDA) bromide (QAD; n=10), and Control (n=6). Heifers were assessed at 0 h, 6, 24, 48, 72 and 168 h post-vaccination; serology was evaluated at first dose (D0), booster (D21) and D42. Heifers vaccinated with Vaccine B (p= 0.0001) and C (p= 0.0001) had a more intense local reaction, while there was a higher rectal temperature detected in heifers vaccinated with Vaccine C (p= 0.020). There was greater systemic reaction observed for heifers vaccinated with Vaccines B and C at 48h (p= 0.002) after a second dose. Clinical pathology parameters [white blood count (WBC) (p = 0.001), neutrophils (p = 0.0001) and haptoglobin concentrations (p = 0.0001)] were higher in animals vaccinated with Vaccine C. Neutralizing Abs against BVDV type 1 strains, NADL and Singer, were detected in animals vaccinated with Vaccines A or C at D42, while BVDV-2 antibodies were detected only in animals vaccinated with Vaccine C. A BHV-1 antibody was detected in all three vaccine groups (Vaccines A, B or C) at day 42 (21 days post booster vaccination). The findings of this research were based on three different commercial laboratory formulations and also according to the conditions which the study was conducted. In this context, vaccine containing mineral oil or Amphigen/QAD presented greater local reactivity and induced a significant systemic inflammatory response. Vaccinated heifers with Alum and Amphigen/QAD commercial vaccines enhanced humoral immune response against BVDV and BHV-1.

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Vaccination is a strategy to the prevention and control of reproductive diseases caused by bovine viral diarrhea virus (BVDV) and bovine herpesvirus type 1 (BoHV-1), however the various compositions of commercial vaccines should be evaluated for their ability to induce protection mediated by antibodies. The objective of this research was to evaluate the production of specific neutralizing Abs against BVDV-1 and 2, and BoHV-1 induced by commercial vaccines composed by different adjuvants. Holstein heifers were vaccinated and distributed in three experimental groups: Group I (G1) was vaccinated with a commercial vaccine containing inactivated BVDV-1, BVDV-2 and BoHV-1 diluted in alum hydroxide as adjuvant (n=9); Group II (G2) was vaccinated with an product containing inactivated strains of BVDV-1, BVDV-2, BoHV-1 and BoHV-5 diluted in oil emulsion as adjuvant (n=10); Group III (G3) was vaccinated with a commercial vaccine containing inactivated BVDV-1 and BVDV-2, besides live modified thermosensitive BoHV-1, diluted in Quil A, amphigen and cholesterol (n=10); A control, non-vaccinated group (n=6) was mock vaccinated with saline. Heifers received two subcutaneous doses of 5mL of each commercial vaccine on the right side of the neck, with 21 days interval. Humoral immune response was assessed by the virus neutralization test (VN) against BVDV-1 (NADL and Singer strains), BVDV-2 (SV253 strain) and BoHV-1 (Los Angeles strain) in serum samples collected on vaccination days zero (D0), 21 (D21) and 42 (D42; 21 days after boosting). Neutralizing Abs against BVDV-1 NADL was detected only in D42, regardless of the vaccine used. Similar geometric mean titers (GMT) for BVDV-1 NADL were observed between G1 (log2=5.1) and G3 (log2=5.1). The seroconversion rate (%) was higher in G1 (78%) when compared to G2 (10%) and G3 (40%). For BVDV-1 Singer, it was also possible to detect Abs production in G1 (log2=5.8, 100% seroconversion rate) and G3 (log2=3.5, seroconversion rate = 60%), only after the booster dose (D42). Neutralizing Abs to BVDV-2 (SV253) were detected only in G3, observing 90% seroconversion associated with high titers of Abs (log2=6.7) after the 2nd dose of vaccine (D42). Heifers from G1 and G3 responded to BoHV-1 after the first dose (D21): G1 (log2=2.5, seroconversion rate = 67%) and G3 (log2=0.7, seroconversion rate = 80%). In D42, a higher magnitude response was observed in the heifers from G3 (log2=6.1, 100%) compared with G1 (log2=4.3, 100%) and G2 (log2=2.7, 60%). Based on the data obtained, it can be concluded that the commercial vaccine contained aluminum hydroxide (G1) was most effective in the induction of antibodies against BVDV-1. On the other hand, this vaccine did not induce the production of neutralizing Abs against BVDV-2. Only the heifers from G3 (Quil A, amphigen and cholesterol) generated neutralizing Abs against BVDV-2. The animals that received commercial vaccine containing oil emulsion as adjuvant (G2) had a weak/undetectable response against BVDV-1 and BVDV-2. The best protective response against BoHV-1 was observed in heifers vaccinated with the live modified thermosensitive virus.

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