Hepatic and splenic immune response during acute vs. chronic Brucella melitensis infection using in situ microscopy

Brucella melitensis is an intracellular bacteria causing disease in humans as an incidental host. The infection initiates as acute flu-like symptoms and may transform into a chronic cyclic infection. This cyclic infection may be partly due to the bacteria’s ability to persist within antigen presenting cells and evade the CD8 + T cell response over long periods of time. This research aims to characterize the immune response of the acute and chronic forms of brucellosis in the murine liver and spleen. We also sought to determine if the exhaustion of the CD8 + T cells was a permanent or temporary change. This was accomplished by using adoptive transfer of acutely infected CD8 + T cells and chronically infected CD8 + T cells into a naïve host followed by re-infection. The histological examination presented supports the concept that exhausted T-cells can regain function through evidence of granulomatous inflammation after virulent challenge in a new host environment.     

Brucella canis in a HIV-infected patient

The true incidence of Brucella canis in humans is unknown, but immunocompromised patients in direct contact with dogs are thought to be at a higher risk for infection. In this case report, we present a human case of B. canis in a patient infected with human immunodeficiency virus.     

Genomic Tandem Repeat Analysis Proves Laboratory‐Acquired Brucellosis in Veterinary (Camel) Diagnostic Laboratory in the United Arab Emirates

We report a case of a 64‐year‐old veterinarian working in a state camel veterinary laboratory who was diagnosed with and treated for acute brucellosis with complicating epididymo‐orchitis. Genomic tandem repeat analysis (MLVA‐16) revealed identical Brucella strains in patient cultures and from different dromedary milk samples positive for Brucella melitensis, thereby confirming the diagnosis of a laboratory acquired infection. The case illustrates the high (airborne) infectivity of brucellosis in laboratory settings and the need to implement vigorous bio‐safety measures in veterinary laboratories handling camel specimen diagnostic veterinary laboratory.     

贵阳市家畜布鲁菌感染情况调查与分析

为了解贵阳市2015年家畜布鲁菌的感染情况,以指导全市家畜布鲁菌病的防控工作,采用虎红平板凝集试验和试管凝集试验对全市8 381份牛、羊、猪血清开展布鲁菌感染情况调查。结果表明,牛、羊和猪布鲁菌病阳性率依次为0.29%(13/4 530)、0.39%(11/2 848)和0%(0/1 003)。说明贵阳市家畜布鲁菌病的流行率在国家控制标准以内,今后宜实行检疫和扑杀相结合的防控策略。     

深度测序技术分析小鼠巨噬细胞感染布鲁菌16M株的转录组学变化

布鲁菌病是由布鲁菌引起的一种人兽共患性传染病,曾被用作生物恐怖战剂。本研究利用深度测序技术对布鲁菌感染小鼠巨噬细胞RAW264．7的转录组学轮廓进行了描述。在感染后4h,筛选出差异表达基因3576个,其中58％的基因表现上调;在感染后24h,筛选出差异表达基因3962个,其中45％的基因表现上调。并且在感染后24h内,变化显著的基因都是与炎症、免疫、吞噬、凋亡紧密相关的。进而我们分析了在感染后24h内显著变化的代谢途径,这些代谢途径包括内质网代谢途径、溶酶体代谢途径、及与凋亡相关的代谢途径;及被显著富集的信号通路,这些信号通路包括凋亡通路、NOD受体信号通路、Fc γR介导的吞噬通路、溶酶体信号通路、p53信号通路、内质网相关蛋白的通路;并且发现B细胞受体和toll样受体信号通路在感染后24h与感染后4h相比被显著富集。本研究建立的巨噬细胞感染布鲁菌后差异表达基因数据库,将为布鲁菌致病机制的逐步阐述奠定基础。     

Multivariate relationships between epidemiologic risk factors and zoonotic infections among military personnel in the country of Georgia: A non‐linear canonical correlation analysis

Zoonotic diseases are endemic in the country of Georgia. Using the non‐linear canonical correlation (NCC) method, the aim of this study was to examine the relationship between thirteen epidemiological risk factors and seropositivity to five zoonotic infections (anthrax, Q fever, tularemia, leptospirosis, and Crimean‐Congo hemorrhagic fever [CCHF]) among Georgian military recruits during 2014–2016. According to this multivariate statistical technique, which is suitable for the analysis of two or more sets of qualitative variables simultaneously, two canonical variables were identified. These variables accounted for 68% of the variation between the two sets of categorical variables (“risk factors” and “zoonotic infections”). For the first canonical variable, there was a relationship among CCHF (canonical loading, which is interpreted in the same way as the Pearson's correlation coefficient, [cl] = 0.715), tick bites (cl = 0.418) and slaughter of animals (cl = 0.351). As for the second canonical variable, Q fever (cl = ?0.604) and leptospirosis (cl = ?0.486) were related to rodents inside and outside home (cl = ?0.346) and sweeping in or around home (cl = ?0.317). The NCC method allows researchers to obtain additional insights into the complex relationship between epidemiological risk factors and multiple zoonotic infections.     

Isolation and sero-genomo-epidemiological studies on Brucella infection in dairy cattle in Meghalaya,India

In this study, we report the serological, bacteriological and whole genome sequencing data of a 6 years study of Brucella abortus in Meghalaya, India. Investigation of 3060 sera samples indicated overall prevalence of 6.4% by Rose Bengal Plate Test and 10.7% by ELISA. Considerably higher prevalence was observed among milk samples (17.5%, n = 362) and in blood samples (37.7%, n = 262) by direct PCR. Clinical samples (n = 94) from late abortion cases yielded 11 B. abortus isolates. Multi-locus sequence typing indicated circulation of single sequence type, ST1. Whole genome sequencing (n = 8) and phylogenomic analysis revealed close clustering of majority of isolates in two clusters alongwith genomes from other countries, indicating global relatedness among B. abortus. Taken together, the results of our study revealed the putative hotspot of infection in the dairy-dominant districts of the state and also calls for concerted One Health based action for prevention and control of this zoonotic disease.     

Pathological changes,distribution and detection of Brucella melitensis in foetuses of experimentally-infected does

BackgroundBrucellosis of goats is caused by Brucella melitensis. It is a re-emerging zoonotic disease in many countries due to transmission from domestic animals and wildlife such as ibex, deer and wild buffaloes.ObjectiveTo describe the pathological changes, identification and distribution of B. melitensis in foetuses of experimentally infected does.MethodsTwelve female goats of approximately 90 days pregnant were divided into 4 groups. Group 1 was exposed intra-conjunctival to 100 µL of sterile PBS while goats of Groups 2, 3 and 4 were similarly exposed to 100 µL of an inoculum containing 10⁹ CFU/mL of live B. melitensis. Goats of these groups were killed at 15, 30 and 60 days post-inoculation, respectively. Foetal fluid and tissues were collected for bacterial identification (using direct bacterial culture, PCR and immuno-peroxidase staining) and histopathological examination.ResultsBilateral intra-conjunctival exposure of pregnant does resulted in in-utero infection of the foetuses. All full-term foetuses of group 4 were either aborted or stillborn, showing petechiations of the skin or absence of hair coat with subcutaneous oedema. The internal organs showed most severe lesions. Immune-peroxidase staining revealed antigen distribution in all organs that became most extensive in group 4. Brucella melitensis was successfully isolated from the stomach content, foetal fluid and various other organs.ConclusionVertical transmission of caprine brucellosis was evident causing mild to moderate lesions in different organs. The samples of choice for isolation and identification of B. melitensis are stomach content as well as liver and spleen tissue.     

Validation of a competitive ELISA for diagnosis of Brucella melitensis infection in sheep and goats

A competitive enzyme-linked immunosorbent assay (cELISA) was validated for the serodiagnosis of Brucella melitensis infection in small ruminants using 2108 positive and 2154 negative reference sera from sheep and goats. The optimum cut-off values, offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp), determined by receiver operating characteristic analysis, were at 23.6%, 21.8% and 25.0% inhibition of the conjugate control for sheep, goats and both species, respectively. The DSns of the cELISA for sheep, goats and both species at these cut-off values were 89.2% (95% confidence interval 87.1-91.1%), 74.0% (95% CI 71.4-76.5%) and 77.9% (95% CI 76.1-79.7%), whereas DSps were 96.4% (95% CI 95.2-97.4%), 92.9% (95% CI 91.1-94.3%) and 97.2% (95% CI 96.4-97.8%), respectively. Compared to cELISA, indirect ELISA and fluorescence polarisation assay have higher DSns and DSps. However, the results obtained with the cELISA were in good agreement with those of the complement fixation test (CFT) under field conditions using 5735 sheep and goat sera. The cELISA can be used as an alternative to the CFT for diagnosing B. melitensis infection in small ruminants.     

Lameness in rams following vaccination against Brucella ovis infection

Extract

Since the introduction in 1957 of a vaccination procedure to control Brucella ovis infection in sheep, many thousands of rams have been vaccinated annually throughout New Zealand almost without complications.     

Brucella melitensis infection in dog: a critical issue in the control of brucellosis in ruminant farms

Canine brucellosis is a contagious disease associated with health implications for humans as well as for a wide range of wild and domesticated animals. In this study, 173 dog blood specimens were sampled from herding dogs in three different provinces including Tehran (n = 127), Qom (n = 40) and Alborz (n = 6) provinces. The presence of Brucella antibodies was determined using Rose Bengal plate test (RBPT), slow agglutination test (SAT) and 2-mercaptoethanol (2-ME), respectively. The seropositive samples were further screened using blood culture and PCR tests to identify and differentiate the implicated Brucella species. According to our results, 24.3% (42/173), 13.8% (24/173) and 6.3% (11/173) of blood samples were tested positive using RBPT, SAT and 2-ME, respectively. However, among 42 seropositive samples, only 38.1% (16/42) and 14.2% (6/42) were positive by PCR and culture, respectively. Brucella melitensis biovar 1 and biovar 2 was isolated from the bacterial cultures of 6 blood samples and confirmed by biotyping, AMOS PCR and Bruce-ladder PCR assays. These findings highlight the potential risk of Brucella transmission from dog to humans along with other livestock and reflect the critical role of infected dogs in the persistence of Brucella infections among ruminant farms. This study stresses the need for further epidemiological investigations on canine brucellosis among herding dogs and suggests the systematic screening of the disease among companion animals such as dogs in order to improve brucellosis surveillance and control programs.     

An ELISA with Brucella lipopolysaccharide antigen for the diagnosis of B. melitensis infection in sheep and for the evaluation of serological responses following subcutaneous or conjunctival B. melitensis strain Rev 1 vaccination.

An indirect enzyme-linked immunosorbent assay (ELISA) with unpurified Brucella melitensis smooth lipopolysaccharide (S-LPS) as antigen was evaluated for the serological diagnosis of B. melitensis infection in sheep in comparison with the Rose Bengal (RB), complement fixation (CF), radial immunodiffusion (RID), microplate agglutination (MA) and rivanol agglutination (RIV) tests. Tests RB and CF detected as positive each of the 77 sera from B. melitensis-infected animals tested, the RID (74), MA (76) and the RIV (72) were less sensitive. However, all tests compared were negative when 77 sera from Brucella-free rams were tested. While subcutaneous Rev 1 vaccination induced high response levels in any of the tests, low level responses were obtained upon conjunctival vaccination, particularly in ELISA and RID tests.     

Imaging diagnosis--Ventriculo-peritoneal shunt associated infection in a dog

Ventriculo-peritoneal shunting is a surgical treatment for hydrocephalus. Complications of this procedure are not well described in dogs. The most common complication in humans is infection, which can be fatal if not diagnosed and treated quickly. We describe the magnetic resonance (MR) imaging characteristics of a shunt-associated cerebral infection in a dog. The MR features of the infection included hyperintensity of the lining of the ventricular system visible on a T2-weighted FLAIR sequence and marked linear contrast enhancement of the ependymal layer on T1-weighted sequences, similar to that described in people.     

Autochthonous Chagas disease in the southern United States: A case report of suspected residential and military exposures

Chagas disease is a parasitic infection that can result in a progressive dilated cardiomyopathy. Here, we present the epidemiologic details of a suspected locally acquired transmission case originating from the southern United States. This is the first published report of Chagas disease in a young, healthy United States veteran with repeat triatomine exposures in Arizona. Military personnel and Arizona residents should be aware of their Chagas disease transmission risks.     

Subclinical paratuberculosis in goats following experimental infection: An immunological and microbiological study

An experimental oral infection of goats with a caprine isolate of Mycobacterium a. subsp. paratuberculosis was used to investigate immunological and bacteriological events during the subclinical phase of infection. Seven goats at 5–8 weeks of age were given a bacterial suspension in milk-replacement three times weekly for 9 weeks. Six animals were kept as controls.

Cellular recall responses against M. a. paratuberculosis were analysed by means of a lymphocyte proliferation test, an IFN-γ assay and an IL-2 receptor assay. All inoculated animals had detectable CMI responses from 9 weeks post-inoculation and through the 2 years of study, although the responses were highest during the first year. Antibodies against M. a. paratuberculosis could be detected from weeks 15–20 in four of the seven animals, and one additional animal became antibody positive at week 35, while two inoculated animals did not produce significant antibody titres during the experiment. At about 1-year post-inoculation, two animals became faecal shedders, while two others started to excrete bacteria into faeces about 2 years post-inoculation. The appearance of M. a. paratuberculosis in faeces was not associated with a decline in cellular responses as far as could be assessed using the current methods for measuring CMI.

Pathological lesions due to M. a. paratuberculosis infection and presence of bacteria were recorded in the intestine and/or mesenteric lymph nodes of five animals while lymph node changes suggestive of paratuberculosis were observed in one animal. Only the two animals with no signs of an active infection at necropsy showed a considerable decline in the cellular parameters during the last year of the study, particularly in the IFN-γ assay.

The two animals with the highest levels of M. a. paratuberculosis responsive CD8+ lymphocytes in the circulation about 1-year post-inoculation had no detectable lesions in the distal ileum and colon at necropsy, while high numbers of γδ T-cells responsive to M. a. paratuberculosis in the circulation were associated with disseminated lesions in the distal ileum and colon.     

Babesia canis vogeli,Ehrlichia canis,and Anaplasma platys infection in a dog

A 12‐month‐old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real‐time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles.     

Risk factors associated with Cryptosporidium parvum infection in cattle

A 2-year, cross-sectional study was conducted to identify risk factors for Cryptosporidium sp. infection in bovine farms in central Italy. Faecal samples were collected on 248 farms, from 2024 calves and analysed using ELISA and immunofluorescent assay (IFA) commercial kits. In all 101 samples confirmed to be positive with IFA, the aetiological agent was identified as Cryptosporidium parvumand a large genetic variability was detected by subtype analysis. The prevalence of farm infection ranged from 3.4% to 35.6%. Univariate analysis showed a number of putative risk factors, including the type of farm, stalling of calves, late supply of colostrum, number of heads and contact between calves and adults. However, multivariate analysis confirmed that the higher risk for calves was associated with housing calves separately from their dams, a characteristic practice of dairy herd, whereas calves being nursed by their dams, a characteristic of cow-calf herd resulted as a protective factor.