Serological and Molecular Survey of Brucella Species in Owners and Their Dogs Living on Island and Mainland Seashore Areas of Brazil.

Background: Although Brucella abortus, Brucella suis, and Brucella canis may infect humans and dogs worldwide, no study to date has assessed and compared owners and their dogs between island and mainland seashore areas. Materials and Methods: Accordingly, the study herein has applied serological tests, including Microplate Agglutination Test with 2-Mercaptoethanol, immunochromatographic assay, and Rose Bengal Test, and a Brucella genus-specific PCR assay to 195 owners and their 148 dogs living on 1 mainland seashore area and three nearby oceanic islands of southern Brazil. Results: No seropositivity to B. abortus and B. suis was detected in owner or dog sera. Anti-B. canis seropositivity was observed in 3/148 (2.0%) dogs, but no owner sample was seropositive to B. canis. In addition, all blood samples from both owners and dogs were negative on Brucella genus-specific PCR assay. Conclusions: The seropositive dogs were not related and lived on the seashore mainland area of Guaraqueçaba city. The absence of seropositivity on the islands and the low seropositivity on the seashore mainland could be attributed to geographic isolation, and suggest the low impact of the disease in the region. Despite being a zoonotic disease, brucellosis by B. canis is not included in the National Program for Control and Eradication of Brucellosis, and its diagnosis and notification are not mandatory. The presence of seropositive dogs highlights the risk to human health and the importance of epidemiological surveillance actions in the region, as well as the need for the implantation of preventive measures to avoid the transmission of the pathogen.

Comparison of Three Serologic Tests for the Detection of Anti-Coxiella burnetii Antibodies in Patients with Q Fever.

The performance of a commercial immunofluorescence assay (IFA commercial), an in-house immunofluorescence assay (IFA in-house) and an indirect enzyme-linked immunosorbent assay (ELISA) were evaluated in the detection of antibodies anti-C. burnetii in the serum of Q fever patients and persons without the disease. For the study, seropositive and seronegative samples for Q fever (n = 200) from a serum bank of the Instituto Adolfo Lutz in Brazil were used. Commercial IFA was considered in this study as the gold standard for diagnosing Q fever. The in-house IFA demonstrated good agreement with the commercial test, showing high sensitivity (91%) and specificity (97%) compared to the gold standard, with a Kappa coefficient of 0.8954. The indirect ELISA test showed lower agreement with the gold standard, showing low sensitivity (67%), although the specificity of the technique was high (97%) and the Kappa coefficient was moderate (0.6631). In-house IFA is an excellent alternative for diagnosing Q fever.

Bacterial identification in cerebrospinal fluid of domestic species with neurologic signs: a retrospective case-series study in 136 animals (2005-2021).

Central nervous system (CNS) infections comprise life-threatening clinical conditions in domestic species, and are commonly related to severe sequelae, disability, or high fatality rates. A set of bacterial pathogens have been identified in central nervous infections in livestock and companion animals, although the most of descriptions are restricted to case reports and a lack of comprehensive studies involving CNS-related bacterial infections have been focused on a great number of domestic species. In this scenario, we retrospectively investigated selected epidemiological data, clinical findings, bacteriological culture, and in vitro susceptibility patterns of 136 nonrepetitive neurologic cases in domestic species (2005-2021). Bacterial isolates were recovered from 25% (34/136) of the cerebrospinal fluid (CSF) sampled. The isolates were obtained from cattle (9/136 = 6.6%), dogs (7/136 = 5.1%), horses (6/136 = 4.4%), goats (3/136 = 2.2%), pigs (3/136 = 2.2%), sheep (3/136 = 2.2%), cats (2/136 = 1.5%), and asinine (1/136 = 0.7%). Among animals with bacterial isolation, Staphylococcus aureus (6/34 = 17.6%), Escherichia coli (5/34 = 14.7%), Staphylococcus beta-hemolytic (5/34 = 14.7%), and Trueperella pyogenes (3/34 = 8.8%) were predominant, in addition to a miscellaneous of other bacteria isolated in minor frequency, e.g., Corynebacterium pseudotuberculosis, Enterobacter cloacae, Mannheimia haemolytica, Pseudomonas aeruginosa, and Streptococcus equi subsp. equi. In vitro susceptibility tests of isolates revealed that amoxicillin/clavulanic acid (11/13 = 84.6%), cephalexin (9/11 = 81.8%), and florfenicol (9/12 = 75%) were the most effective antimicrobials. Conversely, isolates exhibited resistance mainly to tetracycline (6/10 = 60%), penicillin (6/11 = 54.5%), and trimethoprim/sulfamethoxazole (5/11 = 45.5%). Also, multidrug resistance to ≥ 3 classes of antimicrobials was found in 23.5% (8/34) strains. Data relative to the outcome was available in 79.4% (27/34) of animals that had bacterial isolation, and from these, the lethality rate was 92.6% (25/27). Incoordination (14/34 = 41.2%), recumbency (11/34 = 32.4%), apathy (10/34 = 29.4%), anorexia (9/34 = 26.5%), blindness (7/34 = 20.6%), seizure (6/34 = 17.6%), limb paresis (5/34 = 14.7%), head-pressing (4/34 = 11.8%), and nystagmus (3/34 = 8.8%) were the most frequent clinical signs. A variety of bacterial pathogens were identified in the CSF of domestic species showing neurologic signs, with a predominance of staphylococci, streptococci, and enterobacteria. High lethality of cases, poor in vitro efficacy of conventional antimicrobials, and a high in vitro multidrug resistance pattern of isolates were seen. Our results contribute to etiological characterization, antimicrobial resistance patterns, and clinical-epidemiological findings of bacterial infections in domestic species with neurological signs.

Serological Surveillance of Rabies in Free-Range and Captive Common Vampire Bats Desmodus rotundus.

The control of vampire bat rabies (VBR) in Brazil is based on the culling of Desmodus rotundus and the surveillance of outbreaks caused by D. rotundus in cattle and humans in addition to vaccination of susceptible livestock. The detection of anti-rabies antibodies in vampire bats indicates exposure to the rabies virus, and several studies have reported an increase of these antibodies following experimental infection. However, the dynamics of anti-rabies antibodies in natural populations of D. rotundus remains poorly understood. In this study, we took advantage of recent outbreaks of VBR among livestock in the Sao Paulo region of Brazil to test whether seroprevalence in D. rotundus reflects the incidence of rabies in nearby livestock populations. Sixty-four D. rotundus were captured during and after outbreaks from roost located in municipalities belonging to three regions with different incidences of rabies in herbivores. Sixteen seropositive bats were then kept in captivity for up to 120 days, and their antibodies and virus levels were quantified at different time points using the rapid fluorescent focus inhibition test (RFFIT). Antibody titers were associated with the occurrence of ongoing outbreak, with a higher proportion of bats showing titer >0.5 IU/ml in the region with a recent outbreak. However, low titers were still detected in bats from regions reporting the last outbreak of rabies at least 3 years prior to sampling. This study suggests that serological surveillance of rabies in vampire bats can be used as a tool to evaluate risk of outbreaks in at risk populations of cattle and human.

Brucellosis Prevalence in Brazilian Slaughterhouses with Different Meat Inspection Systems.

In Brazil, meat inspection occurs in a decentralized manner and consists of three types: (i) federal inspection (SIF), (ii) state inspection (SISP), and (iii) municipal inspection (SIM). The objective of this work was to discuss the three current inspection systems through the apparent prevalence of bovine brucellosis, a zoonosis that has an eradication program implemented by the Brazilian government. Nine abattoirs from federal, state, and municipal inspection systems were assessed and 1,490 animals were sampled. Serology for brucellosis was determined by the rose bengal test and the complement fixation test. The overall apparent prevalence (and 95% confidence interval) of brucellosis was 2.2% (1.5 to 2.9%). Apparent prevalence stratified by inspection system for SIF, SISP, and SIM was 0.4% (0.0 to 0.9%), 2.0% (0.8 to 3.2%), and 4.3% (2.5 to 6.1%), respectively. Multivariable logistic regression analysis revealed the odds ratio for finding an animal positive for brucellosis among inspection systems. A statistical difference ( P < 0.0015) was observed among surveillance systems, with SISP × SIF, SIM × SISP, and SIM × SIF having an odds ratio of 4,996, 2,304, and 11,494, respectively. Hence, the need for increasing official surveillance in state and municipal inspection systems seems to be necessary and could assist in the surveillance of bovine brucellosis and other diseases of interest to the federation. In addition, an increase in official presence would help to improve the selection of slaughtered cattle during ante- and postmortem inspection, with consequent impact on food safety and public health.

Ticks infesting wild small rodents in three areas of the state of São Paulo, Brazil / Carrapatos infestando pequenos roedores silvestres em três municípios do estado de São Paulo, Brasil

ABSTRACT: From May to September 2011, a total of 138 wild rodents of the Cricetidae family were collected in the cities of Anhembi, Bofete and Torre de Pedra, in São Paulo State. All animals were examined for the presence of ticks, which were collected and identified at species level in the laboratory by morphological examination (for adults, nymphs and larvae) and molecular biology, by sequencing of a fragment of the 16S mitochondrial rDNA (for larvae). A total of 47 ticks (21 larvae and 26 nymphs) were collected from rodents, identified as 21 larvae and 23 nymphs of Amblyomma ovale (collected on Oligoryzomys nigripes, Oligoryzomys flavescens, Sooretamys agouya and Nectomys squamipes), two nymphs of Ixodes schulzei (O. nigripes and O. flavescens) and an Amblyomma sculptum nymph in S. agouya. The overall prevalence of parasitism by A. ovale was 4.3% for larvae, and 10.1% for nymphs. The mean intensity of parasitism was 3.5 larvae/parasitized animal, and 1.6 nymphs/parasitized animal. One O. nigripes was found parasitized by both larva and nymph of A. ovale, and another O. nigripes was parasitized simultaneously by an I. schulzei nymph and five A. ovale nymphs. This research reports the following unpublished records: A. ovale on O. flavescens, O. nigripes and S. agouya; A. sculptum on S. agouya; and I. schulzei on O. flavescens and O. nigripes.

RESUMO: De maio a setembro de 2011, um total de 138 roedores silvestres da família Cricetidae foram coletados nos municípios de Anhembi, Bofete e Torre de Pedra, no estado de São Paulo. Todos os animais foram examinados quanto à presença de carrapatos, os quais foram coletados e identificados ao nível de espécie em laboratório, através de análises morfológicas (para adultos, ninfas e larvas) e por biologia molecular para confirmar estas análises, através do sequenciamento de um fragmento do gene 16S rDNA mitocondrial (para uma parte das larvas). Um total de 47 carrapatos (21 larvas e 26 ninfas) foi coletado dos roedores, sendo identificados como 21 larvas e 23 ninfas de Amblyomma ovale (coletados sobre Oligoryzomys nigripes, Oligoryzomys flavescens, Sooretamys agouya e Nectomys squamipes), duas ninfas de Ixodes schulzei (em O. nigripes e O. flavescens) e uma ninfa de Amblyomma sculptum em S. agouya. A prevalência geral de parasitismo por A. ovale foi de 4,3% e de 10,1% para larvas e ninfas, respectivamente. As intensidades médias de parasitismo foram de 3,5 larvas/animal parasitado e de 1,6 ninfas/animal parasitado. Um O. nigripes foi encontrado parasitado simultaneamente por larva e ninfa de A. ovale, e outro O. nigripes estava parasitado simultaneamente por uma ninfa de I. schulzei e cinco ninfas de A. ovale. O presente trabalho reporta os seguintes registros inéditos: A. ovale em O. flavescens, O. nigripes e S. agouya; A. sculptum em S. agouya; e I. schulzei em O. flavescens e O. nigripes.

Profile of Cytokines and Chemokines Triggered by Wild-Type Strains of Rabies Virus in Mice.

Rabies is a lethal infectious disease that causes 55,000 human deaths per year and is transmitted by various mammalian species, such as dogs and bats. The host immune response is essential for avoiding viral progression and promoting viral clearance. Cytokines and chemokines are crucial in the development of an immediate antiviral response; the rabies virus (RABV) attempts to evade this immune response. The virus's capacity for evasion is correlated with its pathogenicity and the host's inflammatory response, with highly pathogenic strains being the most efficient at hijacking the host's defense mechanisms and thereby decreasing inflammation. The purpose of this study was to evaluate the expression of a set of cytokine and chemokine genes that are related to the immune response in the brains of mice inoculated intramuscularly or intracerebrally with two wild-type strains of RABV, one from dog and the other from vampire bat. The results demonstrated that the gene expression profile is intrinsic to the specific rabies variant. The prompt production of cytokines and chemokines seems to be more important than their levels of expression for surviving a rabies infection.

Evaluation of short-interfering RNAs treatment in experimental rabies due to wild-type virus

ABSTRACTWe have evaluated the efficacy of short-interfering RNAs targeting the nucleoprotein gene and also the brain immune response in treated and non-treated infected mice. Mice were inoculated with wild-type virus, classified as dog (hv2) or vampire bat (hv3) variants and both groups were treated or leaved as controls. No difference was observed in the lethality rate between treated and non-treated groups, although clinical evaluation of hv2 infected mice showed differences in the severity of clinical disease (p = 0.0006). Evaluation of brain immune response 5 days post-inoculation in treated hv2 group showed no difference among the analyzed genes, whereas after 10 days post-inoculation there was increased expression of 2',5'-oligoadenylate synthetase 1, tumor necrosis factor alpha, interleukin 12, interferon gamma, and C-X-C motif chemokine 10 associated with higher expression of Ngene in the same period (p < 0.0001). In hv2 non-treated group only higher interferon beta expression was found at day 5. The observed differences in results of the immune response genes between treated and non-treated groups is not promising as they had neither impact on mortality nor even a reduction in the expression of N gene in siRNA treated animals. This finding suggests that the use of pre-designed siRNA alone may not be useful in rabies treatment.

Evaluation of short-interfering RNAs treatment in experimental rabies due to wild-type virus.

We have evaluated the efficacy of short-interfering RNAs targeting the nucleoprotein gene and also the brain immune response in treated and non-treated infected mice. Mice were inoculated with wild-type virus, classified as dog (hv2) or vampire bat (hv3) variants and both groups were treated or left as controls. No difference was observed in the lethality rate between treated and non-treated groups, although clinical evaluation of hv2 infected mice showed differences in the severity of clinical disease (p=0.0006). Evaluation of brain immune response 5 days post-inoculation in treated hv2 group showed no difference among the analyzed genes, whereas after 10 days post-inoculation there was increased expression of 2',5'-oligoadenylate synthetase 1, tumor necrosis factor alpha, interleukin 12, interferon gamma, and C-X-C motif chemokine 10 associated with higher expression of N gene in the same period (p<0.0001). In hv2 non-treated group only higher interferon beta expression was found at day 5. The observed differences in results of the immune response genes between treated and non-treated groups is not promising as they had neither impact on mortality nor even a reduction in the expression of N gene in siRNA treated animals. This finding suggests that the use of pre-designed siRNA alone may not be useful in rabies treatment.

Outbreak Control and Clinical, Pathological, and Epidemiological Aspects and Molecular Characterization of a Bovine Herpesvirus Type 5 on a Feedlot Farm in São Paulo State.

This paper describes the control, epidemiological, pathological, and molecular aspects of an outbreak of meningoencephalitis in calves due to bovine herpesvirus 5 at a feedlot with 540 animals in São Paulo State, Brazil. The introduction of new animals and contact between the resident animals and the introduced ones were most likely responsible for virus transmission. Bovine herpesvirus 1 vaccine was used, resulting in the efficacy of the outbreak control, although two bovine herpesvirus 1 positive animals, vaccinated and revaccinated, presented meningoencephalitis, thereby characterizing vaccinal failure.

Detecção de Brucella spp. em leite bovino não pasteurizado através da Reação de Cadeia pela Polimerase (PCR) / Detection of Brucella spp. in unpasteurized cow milk by polymerase chain reaction (PCR)

A brucelose é uma importante zoonose causada por bactérias do gênero Brucella. O homem é infectado pelo contato com as secreções reprodutivas como placenta, lóquios placentários, sêmen e secreções penianas de animais infectados ou pelo consumo de leite e derivados não pasteurizados. Com o objetivo de pesquisar a presença da bactéria no leite, foram avaliadas, através da técnica da reação em cadeia da polimerase (PCR), 30 amostras de leite cru comercializadas clandestinamente na região de Botucatu, São Paulo, bem como 50 amostras de leite entregues em laticínio, previamente à pasteurização. Das 80 amostras analisadas pela técnica de PCR, 10 (12,5%) foram positivas e 70 (87,5%) negativas. Dentre as amostras positivas, 5 amostras (16,6%) eram provenientes de comerciantes ilegais e outras 5 amostras (10%) foram obtidas no laticínio. A positividade para Brucella spp. demonstra que o patógeno se encontra presente de forma importante na região de Botucatu, São Paulo, e que o risco associado à saúde pública devido à comercialização de produtos clandestinos sem prévia pasteurização é real.(AU)

Brucellosis is a zoonosis caused by bacteria of the genus Brucella. Man infection occurs through contact with reproductive secretions as placenta and its lochia, semen and penile secretion of infected animals or by consuming unpasteurized milk and dairy products. With the objective of investigating the presence of bacteria in milk, 30 samples of raw milk sold illegally in the region of Botucatu, São Paulo, Brazil, as well as 50 samples of milk delivered to a dairy industry previously to its pasteurization were evaluated by the polymerase chain reaction (PCR) technique. Of the 80 samples analyzed, 10 samples (12.5%) were positive and 70 (87.5%) were negative. Among the positive samples, 5 (16.6%) were from illegal traders and other 5 (10%) were obtained from the dairy industry. Brucella spp. positivity shows that the pathogen is representatively present in Botucatu, São Paulo, Brazil, and the risk associated to public health due to the commercialization of illegal products without pasteurization is real.(AU)

Serological study of vaccinia virus reservoirs in areas with and without official reports of outbreaks in cattle and humans in São Paulo, Brazil.

Vaccinia virus (VACV), the etiological agent of an exanthematic disease, has been associated with several bovine outbreaks in Brazil since the end of the global vaccination campaign against smallpox. It was previously believed that the vaccine virus used for the WHO global campaign had adapted to an unknown wild reservoir and was sporadically re-emerging in outbreaks in cattle and milkers. At present, it is known that Brazilian VACV is phylogenetically different from the vaccinia virus vaccinal strain, but its origin remains unknown. This study assessed the seroprevalence of orthopoxviruses in domestic and wild animals and farmers from 47 farms in three cities in the southwest region of the state of São Paulo with or without official reports of outbreaks in cattle or humans. Our data indicate a low seroprevalence of antibodies in wild animals and raise interesting questions about the real potential of wild rodents and marsupials as VACV reservoirs, suggesting other routes through which VACV can be spread.

Rough virulent strain of Brucella ovis induces pro- and anti-inflammatory cytokines in reproductive tissues in experimentally infected rams.

The ovine brucellosis caused by Brucella ovis has tropism for reproductive tissues but until now the mechanism of bacterial persistence is not understood. Cytokine expression profiles were studied for 8 months in rams after being experimentally infected with the rough virulent strain of B. ovis (R-B. ovis) to study the pathogenesis of B. ovis and immune mechanism possibly associated to bacteria tropism and persistence. The messenger RNA (mRNA) expression levels of interleukin-1α (IL-1α), IL-1ß, IL-6, IL-10, IL-12, interferon-γ (INF-γ) and tumour necrosis factor-α (TNF-α) cytokines were quantified by real-time quantitative RT-PCR (qRT-PCR) in reproductive tissues (epididymus, testicles, ampolae, vesicular glands and bulbourethral glands), and non-reproductive (liver, spleen and kidneys) tissues at 30, 60, 120 and 240 days post infection (dpi). During the acute phase of infection at 30 dpi, the host immune response was most notable demonstrating an up-regulation of several cytokines in reproductive tissues, including the epididymus (IL-6, IL-1ß and IL-1α), testicles (INF-γ and IL-12), bulbourethral glands (IL-6 and TNF-α) and ampolae (INF-γ, IL-10, IL-1ß and IL-1α). During the development of infection, cytokine gene expression levels decreased, providing evidence of immunosuppression and evidence of immune evasion that favoured persistence of chronic R-B. ovis infection. During the chronic phase of R-B. ovis infection (120 and 240 dpi), cytokine production was down-regulated in the epididymus (IL-1ß and IL-1α), testicles (INF-γ and IL-12), and ampolae (INF-γ, IL-10, IL-1ß and IL-1α), with the exception of the bulbourethral glands (IL-6 and TNF-α) and epididymus (IL-6); in these tissues, R-B. ovis infection resulted in up-regulation of the pro-inflammatory cytokine IL-6. Herein, we report cytokine expression profiles in tissues of rams experimentally infected with the rough strain of B. ovis, which are associated with bacterial persistence and macrophage activation.

Vaccinia virus in humans and cattle in southwest region of Sao Paulo state, Brazil.

A new outbreak of Vaccinia virus was observed in Southwest region of São Paulo State, Brazil. The disease was observed in four small dairy farms with manual milking. Lesions were detected in cattle and in humans previously vaccinated and not vaccinated against smallpox. Although several reports of Vaccinia virus outbreaks have been occurring in Brazil, it was not yet reported in this region. This outbreak reinforces the viral circulation in our country. The disease in persons previously vaccinated and not vaccinated against smallpox reinforces the absence of immunity, the risk to the human health, and the need for more epidemiologic and immunologic studies.

Uso de imunomoduladores nas enfermidades infecciosas dos animais domésticos / Use of immunomodulators in infectious diseases of domestic animals

Imunomoduladores são substâncias que atuam no sistema imunológico conferindo aumento da respostaorgânica contra determinados microorganismos, incluindo vírus, bactérias e protozoários, mediante àprodução de interferon e seus indutores. Existem muitas situações na medicina veterinária em que setorna desejável potencializar a resposta imune, principalmente quando se pretende aumentar a resistênciaàs infecções e no tratamento de enfermidades imunossupressoras ou de doenças infecciosas multifatorias,ou seja, nas quais vários agentes estão envolvidos e devido a isso, dificilmente obtêm-se sucesso noemprego de tratamentos convencionais. Na medicina veterinária alguns imunomoduladores utilizadossão interferons , interleucinas, Bacilo de Calmett-Guérin (BCG) e seus derivados, Propionibacteriumacnes (Corynebacterium parvum), vacina bacteriana mista, PIND-ORF, Phosprenyl, Quillaja saponis,Bordetella pertussis, avridina e levamizol. O presente trabalho avalia a aplicação dos diferentesimunomoduladores na profilaxia e na terapêutica das doenças infecciosas dos animais domésticos.

Immunomodulators are substances that act in the immune system providing, increase of the organicanswer against microorganisms, including virus, bacteria and protozoa, by inducing the production ofinterferon and its inducers. There are a lot of situations in veterinary medicine where it is usefull topotencialize the immune response of individuals, mainly when is desired to increase the resistance toinfections and the treatment of immunossupressing or multifactorials infectious diseases. In veterinarymedicine some of more used immunomodulators are interferons and interferon inducers, interleukines,Baccilus of Calmett-Guérin (BCG) and its derivated, Propionibacterium acnes (Corynebacterium parvum),mixed bacterial vaccine, PIND-ORF, Phosprenyl, Quillja Saponis, Bordetella pertussis, avridine and thelevamizole. The present work review the available scientific literature, regarding the use of differentimmunomodulators in the prophylaxis and in the therapeutics of infectious diseases in domestic animals.