Outbreak of Brucellosis in Domestic Elk in Korea

Seven of 18 elk on a deer farm were found by the official Rose‐Bengal agglutination test (RBT) and tube agglutination test to be brucellosis reactors/suspects. Evaluation with the competitive ELISA (C‐ELISA) and the fluorescence polarization assay (FPA) tests revealed that six and five sera were positive respectively. The seven reactors/ suspects were slaughtered and their blood and tissues were collected. Brucella species could be isolated from three of the slaughtered animals, with nine isolates being obtained from the popliteal, supramammary and submandibular lymph nodes, vaginal discharge, mammary tissue and spleen. Brucella genus‐specific PCR based on 16S rRNA and AMOS‐PCR, which is specific for differential Brucella species, revealed that all nine isolates were Brucella abortus. These nine were further confirmed to be B. abortus biovar 1 by classical biotyping scheme assays. This is the first report of an outbreak of brucellosis in domestic elk in Korea. Our observations suggest that deer should be included in the routine Brucella surveillance programme for the effective control and prevention of brucellosis in Korea.     

Brucella-associated cervical bursitis in cattle

Bovine brucellosis poses a risk to human health and causes serious economic losses for the animal industry. This report describes the use of different diagnostic methods for the diagnosis of brucellosis in cattle affected by cervical bursitis from a slaughterhouse located in São Luís, Maranhão, Brazil. Serum samples from a total of 47 cattle with bursitis were collected and submitted to the Rose Bengal Test (RBT), and RBT-positive samples were further confirmed by the 2-mercaptoethanol (2-ME) assay. RBT indicated 85.1% (40/47) of positive samples, from which 78.7% (37/47) were confirmed by 2-ME. Immunohistochemistry detected Brucella spp. in 34.0% (16/47) of tissues with bursitis. PCR and/or bacterial isolation demonstrated that 63.8% (30/47) of samples were positive and morphologically compatible with Brucella sp. All colonies suggestive of Brucella sp. were confirmed by PCR. Isolates were further characterized by PCR Multiplex AMOS-ENHANCED, which indicated that the isolates corresponded to biovar 1, 2, 4 (43.33%). This study evidences an association between cervical bursitis and Brucella spp. infection in cattle, and that different biovars of Brucella circulate in bovine herds in Maranhão.

     

Brucella abortus is Prevalent in Both Humans and Animals in Bangladesh

To determine the role of different Brucella (B.) spp. in Bangladesh, 62 animal samples and 500 human sera were tested. Animal samples from cattle, goats and sheep (including milk, bull semen, vaginal swabs and placentas) were cultured for Brucella spp. Three test‐positive human sera and all animal samples were screened by Brucella genus‐specific real‐time PCR (RT‐PCR), and positive samples were then tested by IS711 RT‐PCR to detect B. abortus and B. melitensis DNA. Only B. abortus DNA was amplified from 13 human and six animal samples. This is the first report describing B. abortus as the aetiological agent of brucellosis in occupationally exposed humans in Bangladesh. Of note is failure to detect B. melitensis DNA, the species most often associated with human brucellosis worldwide. Further studies are required to explore the occurrence of Brucella melitensis in Bangladesh.     

Risk factors associated with camel brucellosis in Jordan

During the period between February, 2004 and December, 2006, a cross-sectional study was performed to investigate some epidemiological aspects related to camel brucellosis in Jordan. Four hundred twelve camel sera from 37 herds were randomly collected and analyzed using Rose Bengal plate test and complement fixation test. A structured pre-tested questionnaire was administered to collect information on camel herd health and management. A multivariable logistic regression model was constructed to investigate risk factors associated with seropositivity to Brucella antigens. Moreover, the incidence of Brucella-specific abortion was investigated in 7 camel herds located in different locations in Southern Jordan. The true prevalence of Brucella-seropositive in camels was 12.1%. Thirteen (35.1%) herds had at least one positive camel. The seroprevalence of brucellosis in camels was significantly higher in the southern part of Jordan than that in central or northern Jordan. The multivariable logistic regression model on both individual and herd levels revealed large herds and contact with small ruminants as risk factors for Brucella seropositivity. On the other hand, using disinfectants was identified as a protective factor (OR = 0.8; 95% CI: 0.1, 0.9) only on the camel herd level. The incidence of Brucella-caused abortion was 1.9%. Brucella melitensis biotype 3 was isolated from 4 aborted camel fetuses.     

A comparison of the potency of several Brucella allergens used to detect brucellosis in cattle

The potency of Brucella allergens prepared from a smooth Brucella abortus strain S-99, mucoid strain Leewarden, rough strain 45/20, and rough Brucella melitensis strain B-115 was assessed. The potency of these allergens was compared with that of a standard allergen prepared from smooth Brucella abortus S-99 that efficiently detected bovine brucellosis in other studies. Eight cattle experimentally inoculated with Brucella abortus 544 were tested with the allergens 4 and 10 weeks after infection, and again 8 months after infection. All the allergens effectively detected infection but there was a clear distinction in the mean skin reactions 48 and 72 h after injection of the allergens. The skin reactions provoked by the allergens prepared from smooth or mucoid strains of Brucella were most pronounced 48 h after injection. Skin reactions provoked by allergens prepared from rough strains of Brucella were strongest 72 h after injection. Allergens prepared from smooth or mucoid Brucella strains were more potent in detecting brucellosis than those prepared from rough strains of Brucella.Abbreviations Bruc/OCB Brucellergen OCB - cfu colony-forming units - CFT complement fixation test - ID-DLO Institute voor Dierhouderij en Diergezondheid-Dienst Landbouwkundig Onderzoek - ICFTU international complement fixation units - IU international units - LPS lipopolysaccharide - SAT serum agglutination test - SDTH skin delayed-type hypersensitivity     

Fatal Pneumonia and Pleuritis Caused by an Agujon Beak Penetration in a Bottlenose Dolphin from Puerto Rico

Abstract

Although wounds caused by fish spines have been reported to cause disease and mortality in marine mammals, no record of this phenomenon exists for the Caribbean area. We report a case of disease and consequent mortality in a bottlenose dolphin Tursiops truncatus caused by ingestion of the beak of an agujon Tylosurus acus in Manatí, Puerto Rico. Internal examination of the dolphin revealed that its left lung was necrotic and reduced to one-fourth its normal size and that it had chronic–active pleural inflammation. A 9.1-cm compound bone fragment was found embedded in an abscess in the pleural lining of the left lung. The bone fragment was identified as belonging to the jaws of an agujon, a circumtropical, polytypic species commonly found in the Caribbean Sea. Histopathology revealed chronic inflammation and fibrosis of the lung in addition to granulomatous masses in the pleural space. The cause of death was determined to be chronic lung infection caused by migration of the agujon's beak from the dolphin's esophagus or stomach. No similar cases have been observed in the more than 150 marine mammal strandings documented in Puerto Rico and the Virgin Islands since 1985. This is the first report of disease and death caused by migration of an agujon beak in a marine mammal.     

Brucellosis among smallholder cattle farmers in Zambia

A cross-sectional study was performed in Southern and Lusaka provinces of Zambia between March and September 2008 to estimate Brucella seroprevalence in cattle kept by smallholder dairy farmers (n = 185). Rose Bengal test (RBT) was used as a screening test followed by confirmation with competitive ELISA (c-ELISA). We investigated 1,323 cattle, of which 383 had a history of receiving vaccination against brucellosis and 36 had a history of abortion. Overall seroprevalence was 6.0% with areas where vaccination was practiced having low seroprevalence. Age was associated with Brucella seropositivity (P = 0.03) unlike cattle breed (P = 0.21) and sex (P = 0.32). At area level, there was a negative correlation (Corr. coeff = −0.74) between percentage of animals with brucellosis vaccination history (vaccination coverage) and level of brucellosis; percentage of animals with history of abortion (Corr. coeff. = −0.82) and brucellosis vaccination coverage. However, a positive correlation existed between brucellosis infection levels with percentage of animals having a history of abortion (Corr. coeff. = 0.72). History of vaccination against brucellosis was positively associated with a positive Brucella result on RBT (P = 0.004) whereby animals with history of vaccination against brucellosis were more likely to give a positive RBT test results (OR = 1.52). However, the results of c-ELISA were independent of history of Brucella vaccination (P = 0.149) but was positively associated with history of abortion (OR = 4.12). Our results indicate a relatively low Brucella seroprevalence in cattle from smallholder dairy farmers and that vaccination was effective in reducing cases of Brucella infections and Brucella-related abortions. Human exposure to Brucella through milk from smallholder farmers could result through milk traded on the informal market since that milk is not processed and there no quality and safety controls.     

Diagnosis of Canine Brucellosis: Comparison between Serological and Microbiological Tests and a PCR Based on Primers to 16S-23S rDNA Interspacer

A pair of primers directed to 16S-23S rDNA interspacer (ITS) was designed directed to Brucella genetic sequences in order to develop a polymerase chain reaction (PCR) putatively capable of amplifying DNA from any Brucella species. Nucleic acid extracts from whole-blood from naive dogs were spiked with decreasing amounts of Brucella canis RM6/66 DNA and the resulting solutions were tested by PCR. In addition, the ability of PCR to amplify Brucella spp. genetic sequences from naturally infected dogs was evaluated using 210 whole-blood samples of dogs from 19 kennels. The whole-blood samples collected were subjected to blood culture and PCR. Serodiagnosis was performed using the rapid slide agglutination test with and without 2-mercaptoethanol. The DNA from whole blood was extracted using proteinase-K, sodium dodecyl sulphate and cetyl trimethyl ammonium bromide followed by phenol–chloroform purification. The PCR was capable of detecting as little as 3.8 fg of Brucella DNA mixed with 450 ng of host DNA. Theoretically, 3.8 fg of Brucella DNA represents the total genomic mass of fewer than two bacterial cells. The PCR diagnostic sensitivity and specificity were 100%. From the results observed in the present study, we conclude that PCR could be used as confirmatory test for diagnosis of B. canis infection.     

Investigation of bovine brucellosis in the Northeastern Turkey

Bovine brucellosis, caused by Brucella abortus, is a significant problem for both public and animal health in Turkey. This study was conducted on the calving seasons between 2001 and 2006. A total of 626 serum samples of cattle obtained from 27 herds with a history of abortions was examined for Brucella antibodies by RBPT, SAT and ELISA. Of the cattle sera analysed, 221 (35,30%) and 206 (32, 92%) and 247 (39,45%) were found to be positive by RBPT , SAT and ELISA, respectively. B. abortus was isolated from 48 (32,21%) of 149 lung samples and stomach contents of the aborted fetuses. Based on the biochemical tests and the agglutination tests with monospecific A and M antisera, only 3 of the isolates were found to be B. abortus biotype 1 and the remaining 45 were biotype 3. This study also revealed that the dominant biotype of B. abortus was biotype 3 in this region. The determination of the agents responsible for bovine brucellosis and serosurvey of this disease are expected to help better understanding of this zoonotic infection in this region and neighbouring countries.     

Prevalence and risk factors associated with bovine brucellosis in herds under extensive production system in southwestern Nigeria

Using a cross-sectional survey, we determined the prevalence and risk factors associated with bovine brucellosis in herds under extensive production system in southwestern Nigeria. Antibodies to Brucella species in serum samples were tested using the Rose Bengal test (RBT) and competitive enzyme-linked immunosorbent assay (cELISA); for milk, the milk ring test (MRT) and indirect-ELISA (i-ELISA) were used. Questionnaire was administered to cattle herdsmen to determine factors predisposing the animals to bovine brucellosis. Data were analyzed using STATA 12. From 513 serum and 635 milk samples tested among 120 herds, overall animal-level prevalence of 10.1% (95% CI 7.5–12.7%) and 20.2% (95% CI 17.1–23.3%) were recorded by RBT and MRT, respectively; while 9.4% (95% CI 6.9–11.9%) and 17.8% (95% CI 14.8–20.8%) were obtained using cELISA and i-ELISA, respectively. In all, from the 120 herds tested, 29.2% and 43.3% were positive by RBT and MRT, respectively. Multivariable logistic regression revealed that herd location (OR?=?8.12, 95% CI 1.68–38.90) and improper disposal of placenta/fetus (OR?=?17.33, 95% CI 4.81–62.33) were predictors for a seropositive herd using RBT; while herd location (OR?=?5.13, 95% CI 1.27–20.28), large herd size (OR?=?2.62, 95% CI 1.15–5.85), and occurrence of abortion for a year or more (OR?=?4.62, 95% CI 1.53–13.71) were predictors of seropositivity to antibodies to Brucella spp. using MRT. We found high prevalence of brucellosis in cattle herds under extensive management system in southwestern Nigeria. Urgent and coordinated control strategies are required to mitigate this problem.

     

A review of Brucella sp. infection of sea mammals with particular emphasis on isolates from Scotland

Brucellae recovered from sea mammals were first reported in 1994. In the years since both culture and serological analysis have demonstrated that the infection occurs in a wide range of species of marine mammals inhabiting a vast amount of the world’s oceans. Molecular studies have demonstrated that the isolates differ from those found amongst terrestrial animals and also distinguish between strains which have seals and cetaceans as their preferred hosts. At the phenotypic level seal and cetacean strains can also be differed with respect to their CO₂ requirement, primary growth on Farrells medium and metabolic activity on galactose. Two new species B. cetaceae and B. pinnipediae have been proposed as a result. This paper provides a review of Brucella in sea mammals and updates findings from the study of sea mammals from around the coast of Scotland.     

Seroprevalence of brucellosis and its contribution to abortion in cattle,camel, and goat kept under pastoral management in Borana,Ethiopia

The involvement of Brucella infection in causing abortion was investigated in a breeding female subpopulation of 283 cattle, 756 camels, and 757 goats. Serum samples were serially tested using the Rose Bengal test and complement fixation test. The study showed that anti-Brucella antibodies were prevalent in 10.6% (95% confidence interval (CI), 7.4, 14.9), 2.2% (95%CI, 1.4, 3.7), and 1.9% (95%CI, 1.1, 3.2) of cattle, camel, and goats, respectively. Abortion was more commonly reported in camels (23.4%) than cattle (13.8%) and goats (12.4%). The results of this study suggested that Brucella infections contribute significantly to abortion in cattle (odds ratio (OR), = 4.7; 95%CI, 2.0, 10.8) and goats (OR = 6.9; 95%CI, 2.2, 21.7) but not in camels. The number of young animals produced by breeding females seems to be apparently reduced in seropositive groups. Keeping more than two animal species at household level was found to be the risk factor for cattle (OR = 3.1; 95%CI, 1.2, 7.9) and camel (OR = 5.3; 95%CI, 1.2–23.5) seropositivity to Brucella infection when compared to those animals from households that keep only two animal species. This may suggest a possibility of cross species transmission of Brucella infection under such mixed herding. Wet season (OR = 4.8; 95%CI, 1.3, 18.1) was found to be associated with seropositivity in goats, linked to a coincidence of increased deliveries in flocks with possible excretion of Brucella organisms. The study results suggest that Brucella infection is the likely cause of abortion in cattle and goats while other causes largely outweigh brucellosis as a cause of abortion in camels in Borana, hence, contributing to reproductive loss.     

Serological survey of pre-weaned New Zealand fur seals (Arctocephalus forsteri) for brucellosis and leptospirosis

AIM: To conduct a longitudinal serological survey for evidence of Brucella spp and Leptospira spp infection of pre-weaned New Zealand fur seals in a colony on the Otago Peninsula.

METHODS: Seal pups were repeatedly captured on a monthly basis from February through to July 2001. Pups were tagged at first capture and a blood sample was taken at each capture event. A total of 163 sera were collected from 118 seal pups. Where sufficient volume was collected, the sera were tested for leptospirosis using the microscopic agglutination test (MAT), and for brucellosis using the competitive enzyme-linked immunosorbent assay (ELISA) for Brucella abortus.

RESULTS: None of 128 sera from 101 seals tested positive to the ELISA for B. abortus. All tests for Leptospira interrogans serovars Grippotyphosa, Copenhageni, Bratislava and Leptospira borgpetersenii serovar Ballum were negative at a cut-off of <1/100 dilution. Positive or suspicious titres were found to L. interrogans serovars Canicola and Pomona and L. borgpetersenii serovar Hardjo. The highest titres (12,800) were found to serovar Pomona. The titre to serovar Pomona in one seal rose from <1/50 in March to 12,800 in April and was <1/50 when re-sampled in July. The titre to serovar Pomona in another seal dropped from 12,800 in May to <1/50 in June. These seals also had titres to serovar Hardjo, which rose or fell in the same manner. All suspicious or positive titres occurred in late April and early May, when the pups were approximately 4–5 months old. In June and July, all seals tested were negative.

CONCLUSIONS: There was no serological evidence of Brucella infection in the pre-weaned fur seals at the colony. Positive titres to serovars Pomona, Hardjo, or Canicola suggest that a Leptospira species was present at the colony, however isolation or visualisation of the organism is required to confirm this. Care should be exercised when handling New Zealand fur seals to prevent human infection or inadvertent transfer of leptospirosis to another marine mammal species.     

Prevalence of antibodies to Brucella spp. and individual risk Factors of Infection in Traditional Cattle, Goats and Sheep Reared in Livestock–Wildlife Interface Areas of Zambia

A cross-sectional study was performed in the livestock–wildlife interface areas of Lochinvar and Blue Lagoon National Parks and the non-interface area of Kazungula to determine the prevalence of antibodies to Brucella spp. in domestic ruminants and identify individual animal risk factors of infection. A total of 1245 cattle from 124 herds and 280 goats and sheep from 29 flocks were tested sequentially for Brucella antibodies using the Rose Bengal test (RBT) and competitive ELISA. In cattle, individual seroprevalence ranged from 14.1% to 28.1%, while herd sero–prevalence ranged from 46.2% to 74.0% in the three study areas. No goat or sheep tested positive for Brucella antibodies. Three types of cattle grazing strategies were encountered: locally grazed herds (LGH), transhumantly grazed herds (TGH) and river flood plain grazed herds (FGH). Brucella seroprevalence was seen to vary according to area and grazing strategy: Lochinvar and transhumant grazed herds recorded the highest figures, respectively. Age, sex and history of abortion were found to have independent effects on individual seroprevalence. This study establishes that brucellosis is endemic in domestic animals in the livestock–wildlife interface areas of Blue Lagoon and Lochinvar national parks and the disease is also present in Kazungula. We observed that type of grazing strategy had significant impact on cattle Brucella seroprevalence and that transhumant herds were at high risk of being infected.     

Assay dependence of Brucella antibody prevalence in a declining Alaskan harbor seal (Phoca vitulina) population

Background

Brucella is a group of bacteria that causes brucellosis, which can affect population health and reproductive success in many marine mammals. We investigated the serological prevalence of antibodies against Brucella bacteria in a declining harbor seal population in Glacier Bay National Park, Alaska.

Results

Prevalence ranged from 16 to 74 percent for those tests detecting antibodies, indicating that harbor seals in Glacier Bay have been exposed to Brucella bacteria. However, the actual level of serological prevalence could not be determined because results were strongly assay-dependent.

Conclusions

This study reinforces the need to carefully consider assay choice when comparing different studies on the prevalence of anti–Brucella antibodies in pinnipeds and further highlights the need for species- or taxon-specific assay validation for both pathogen and host species.     

Investigation of abortions in brucellosis tested herds

The mandatory reporting and investigation of bovine abortions provides a system for the surveillance of the progress of the Brucellosis Eradication Scheme and a warning of resurgence of brucellosis in tested herds. Before the scheme was introduced in 1971, about a quarter of all abortions were caused by Brucella abortus. By 1976, the percentage of abortions due to Brucella in herds under test for brucellosis had been reduced to 3.4%. Most Brucella abortions occurred during the seventh and eighth months of gestation. Cows in the 4- to 6-year-old age group were at greater risk of aborting from Brucella infection than heifers. Approximately 94% of all aborting cows were in herds which experienced three or fewer abortions. Less than 4% of herds experienced four or more abortions annually. In most cases of abortion due to brucellosis, more than one diagnostic test yielded positive results. However, 13.3% of cases were recognised by positive bacteriological results alone.     

Meta-analysis of Brucella seroprevalence in dairy cattle of Ethiopia

This meta-analysis estimates a single-group summary (effect size) for seroprevalence of Brucella spp. exposure in dairy cattle of Ethiopia. It also attempts to identify study-level variables that could explain the variation in apparent seroprevalence. The literature search was restricted to studies published in English language from January 2000 to December 2013. A template was designed to retrieve the most biologically plausible and consistent variables from the articles. A total of 29 published papers containing 40 animal-level studies were used in the analyses. The single-group summary of Brucella seroprevalence in cattle was estimated to reach 3.3 % with 95 % confidence interval (CI) (2.6–4.2 %). Of all the variables considered, region was the only specific factor identified to explain about 20 % of between-study variation. Accordingly, the region-based meta-analysis forest plot revealed the highest prevalence in central Ethiopia followed by southern part. The lowest prevalence estimate was observed in the western part of the country. The visual inspection of the funnel plot demonstrated the presence of possible publication bias which might dictate shortage of studies with higher prevalences or variance inflation due to infectiousness of Brucella. In conclusion, the quantitative review showed the seroprevalence to be low but widely distributed. More importantly, the review underscores the need for isolation and characterization of the circulating Brucella spp. to capture the type of Brucella spp. involved and its distribution in cattle in Ethiopia.     

Adenovirus and mycoplasma infection in an ornate box turtle (Terrapene ornata ornata) in Hungary

A female, adult ornate box turtle (Terrapene ornata ornata) with fatty liver was submitted for virologic examination in Hungary. Signs of an adenovirus infection including degeneration of the liver cells, enlarged nuclei and intranuclear inclusion bodies were detected by light microscopic examination. The presence of an adenovirus was later confirmed by obtaining partial sequence data from the adenoviral DNA-dependent DNA-polymerase. Phylogenetic analyses revealed that this novel chelonian adenovirus was distinct from previously described reptilian adenoviruses, not belonging to any of the recognized genera of the family Adenoviridae. As a part of the routine diagnostic procedure for chelonians the detection of herpes-, rana- and iridoviruses together with Mycoplasma spp. was attempted. Amplicons were generated by a general mycoplasma polymerase chain reaction (PCR) targeting the 16S/23S ribosomal RNA (rRNA) intergenic spacer region, as well as, a specific Mycoplasma agassizii PCR targeting the 16S rRNA gene. Based on the analyses of partial sequences of the 16S rRNA gene, the Mycoplasma sp. of the ornate box turtle seemed to be identical with the recently described eastern box turtle (Terrapene carolina carolina) Mycoplasma sp. This is the first report of a novel chelonian adenovirus and a mycoplasma infection in an ornate box turtle (T. ornata ornata) in Europe.     

Ophthalmomyiasis interna anterior in a dog: keratotomy and extraction of a Cuterebra sp. larva

A 1‐year‐old female spayed mixed‐breed dog was presented for evaluation of acute onset of lethargy and blepharospasm OD. Slit‐lamp biomicroscopy revealed a segmented larva embedded in the inferonasal iris. Moderate anterior uveitis was present. After pharmacologic dilation, mild vitreal hemorrhage and chorioretinal migration tracks were identified. The dog was treated empirically for uveitis for 5 days, after which a keratotomy and larval foreign body extraction was performed. The dog recovered uneventfully from intraocular surgery and remained visual and comfortable. Parasite evaluation confirmed the larva to be a Cuterebra sp. To the authors’ knowledge, this is the first report of canine ophthalmomyiasis interna anterior, and the first report of successful surgical removal from the anterior segment in a dog with preservation of vision.     

Identification of Brucella spp. in feral swine (Sus scrofa) at abattoirs in Texas,USA

Various tissues, nasal swabs, urine and blood samples were collected from 376 feral swine at two federally inspected abattoirs in Texas during six separate sampling periods in 2015. Samples were tested for Brucella spp. by culture and serology. Brucella spp. were cultured from 13.0% of feral swine, and antibodies were detected in 9.8%. Only 32.7% of culture‐positive feral swine were also antibody positive, and 43.2% of antibody‐positive feral swine were culture positive. Approximately, the same number of males (14.0%) and females (12.1%) were culture positive, and slightly more males (10.5%) than females (8.7%) were antibody positive. Our results indicate that serology likely underestimates the prevalence of feral swine infected, and that those who come in contact with feral swine should be aware of the symptoms of infection with Brucella spp. to ensure prompt treatment.