Densitometric analysis of rep-PCR data: Insight into genetic variability and transmission of Clostridium perfringens typed with an improved multiplex PCR.

An epidemiological study was conducted in North-East India (part of Indo-Burma biodiversity hotspot) to better understand the distribution, diversity, and transmission of Clostridium perfringens among livestock, pets, wild animals (captive), and humans. A total of 160 C. perfringens isolates were recovered from 642 diarrhoeic faecal samples with an isolation rate of 24.92%. Isolation rate was the highest among captive wild animals (37.5%) followed by dog (34.6%), human (33.8%), pig (32.7%), cattle (20.8%), goat (18.3%) and poultry (9.3%). Isolates were toxin typed using a seven gene multiplex PCR designed for simultaneous detection of cpa, cpb, cpb2, etx, iap, cpe and netB. The majority of isolates, 128 (80%) were of type A, followed by 17 (10.62%), 5 (3.12%), 4 (2.5%), 3 (1.87%), 2 (1.25%) and 1 (0.63%) isolates of type C, D, E, G, F and B, respectively. Beta 2 toxin gene was present in 65 (50%) of type A isolates, followed by 7 (41.2%), 4 (80%), 1(25%), and 1 (100%) of type C, D, G and B isolates, respectively. Beta 2 toxin has a high prevalence among dogs (28.6%), cattle (27.3%), and pig (20.8%) compared to humans, goat, wild animals, and poultry (1.2-14.3%). The prevalence of CPE and NetB toxin-positive strains was low, with only 3 (1.8%) and 5 (3.1%) isolates, respectively. Association of C. perfringens with diarrhoea in Civet Cat, Golden Langur, and Gray Langur has been reported for the first time. The genetic diversity and transmission of isolates were investigated using automated rep-PCR (Diversilab®, bioMérieux) using two densitometry-based matrices: modified Kullback-Leibler (KL) and Pearson's correlation (PC). The PC and modified KL matrices formed three distinct clusters with 59% and 27.2% similarity, respectively. C. perfringens diversity and transmission were best studied using modified KL matrix that placed more emphasis on the presence of bands rather than intensity. However, the PC method was found to be more suitable for differentiating strains within a toxin type, with slightly higher D-values.

Peste des petits ruminants in Africa: a review of currently available molecular epidemiological data, 2020.

Small ruminants (e.g., sheep and goats) contribute considerably to the cash income and nutrition of small farmers in most countries in Africa and Asia. Their husbandry is threatened by the highly infectious transboundary viral disease peste des petits ruminants (PPR) caused by peste-des-petits-ruminants virus (PPRV). Given its social and economic impact, PPR is presently being targeted by international organizations for global eradication by 2030. Since its first description in Côte d'Ivoire in 1942, and particularly over the last 10 years, a large amount of molecular epidemiological data on the virus have been generated in Africa. This review aims to consolidate these data in order to have a clearer picture of the current PPR situation in Africa, which will, in turn, assist authorities in global eradication attempts.

The elephant-livestock interface modulates anthrax suitability in India.

Anthrax is a potentially life-threatening bacterial disease that can spread between wild and livestock animals and humans. Transmission typically occurs indirectly via environmental exposure, with devastating consequences for human and animal health, as well as pastoralist economies. India has a high annual occurrence of anthrax in some regions, but a country-wide delineation of risk has not yet been undertaken. The current study modelled the geographical suitability of anthrax across India and its associated environmental features using a biogeographic application of machine learning. Both biotic and abiotic features contributed to risk across multiple scales of influence. The elephant-livestock interface was the dominant feature in delineating anthrax suitability. In addition, water-soil balance, soil chemistry and historical forest loss were also influential. These findings suggest that the elephant-livestock interface plays an important role in the cycling of anthrax in India. Livestock prevention efforts targeting this interface, particularly within anthropogenic ecotones, may yield successes in reducing ongoing transmission between animal hosts and subsequent zoonotic transmission to humans.

Evaluation of the immunogenicity and efficacy of BCG and MTBVAC vaccines using a natural transmission model of tuberculosis.

Effective vaccines against tuberculosis (TB) are needed in order to prevent TB transmission in human and animal populations. Evaluation of TB vaccines may be facilitated by using reliable animal models that mimic host pathophysiology and natural transmission of the disease as closely as possible. In this study, we evaluated the immunogenicity and efficacy of two attenuated vaccines, BCG and MTBVAC, after each was given to 17 goats (2 months old) and then exposed for 9 months to goats infected with M. caprae. In general, MTBVAC-vaccinated goats showed higher interferon-gamma release than BCG vaccinated goats in response to bovine protein purified derivative and ESAT-6/CFP-10 antigens and the response was significantly higher than that observed in the control group until challenge. All animals showed lesions consistent with TB at the end of the study. Goats that received either vaccine showed significantly lower scores for pulmonary lymph nodes and total lesions than unvaccinated controls. Both MTBVAC and BCG vaccines proved to be immunogenic and effective in reducing severity of TB pathology caused by M. caprae. Our model system of natural TB transmission may be useful for evaluating and optimizing vaccines.

Transmission dynamics of foot and mouth disease in selected outbreak areas of northwest Ethiopia.

Foot and mouth disease (FMD) is a highly contagious and economically important disease of cloven-hoofed animals, which is endemic in Ethiopia. An outbreak follow-up study was undertaken to quantify the transmission parameters of FMD in the crop-livestock mixed (CLM) system and commercial dairy farms in selected areas of northwest Ethiopia. The transmission parameters were quantified using a generalised linear model (GLM) based on a susceptible-infectious-recovered (SIR) epidemic model. The per day average transmission rate between animals was 0.26 (95% CI 0.22-0.32) and 0.33 (95% CI 0.21-0.57) in the CLM system and in the commercial dairy farms, respectively. The average basic reproduction ratio of FMD was 1.68 (95% CI 1.42-2.07) in the CLM system and 1.98 (95% CI 1.26-3.42) in the commercial dairy farms. The medium per day transmission rate and moderate basic reproduction ratio observed in this study indicated that a vaccination coverage needed to stop transmission of the disease in these populations might not be very high.

Experimental infection of Korean native goats (Capra aegagrus hircus) with bovine viral diarrhea virus 1b.

BACKGROUND: Bovine viral diarrhea virus (BVDV) infects various ungulates and causes reproductive failure in infected goats. BVDV has been detected among goats in the Republic of Korea, but the route of transmission remains unclear. Here, we aimed to investigate whether BVDV-1b circulating among Korean cattle can be transmitted to Korean native goats (Capra aegagrus hircus) and characterize the outcomes of BVDV infection in these goats. RESULTS: Four goats were inoculated intranasally with the Korean noncytopathic (ncp) BVDV-1b strain. Two goats exhibited clinical signs of illness, including coughing and nasal discharge. Nasal swabs and blood were collected to screen for viral RNA and BVDV antibodies. Using the 5'-untranslated region (UTR), viral RNA was detected in the nasal swabs of two goats (Goat 1 and 3) on 12 day post-inoculation (dpi) and in the blood sample of one goat (Goat 1) on 7 and 19 dpi. Using the N-terminal protease (Npro) region, viral RNA was detected in the blood sample of Goat 1 on 7 and 12 dpi. Antibodies to BVDV were detected in Goats 1 and 3 on 16-21 dpi using enzyme-linked immunosorbent assay. Sequence analysis of the virus from nasal swabs and blood samples, which was detected via RT-PCR, using the 5'-UTR and Npro regions led to the identification of the strain as ncp BVDV-1b and revealed changes in the nucleotide sequence of these goats. CONCLUSIONS: Our results indicate that changes in the nucleotide sequence are associated with the establishment of BVDV infection in Korean native goats; these changes may be owing to a process required for the establishment of infection in a new host reservoir. Broadly, these findings highlight the importance of BVDV surveillance in ungulates other than cattle.

Spread and impact of goat pox ("sagolay bohonta") in a village smallholder community around Kaziranga National Park, Assam, India.

During September and October 2017, a highly fatal outbreak of a disease clinically indistinguishable from goat pox occurred in the villages around the Kaziranga National Park, Assam, India. This was investigated through clinical examination of affected animals, individual interviews with goat keepers and participatory village meetings. Laboratory confirmation was impractical due to the isolation and poverty of the affected community and unnecessary due to the specific nature of the clinical signs. Respondents reported not having encountered the disease previously, and it would appear that a naïve local population developed within an endemically affected region because of a trend to avoid purchasing animals from outside the village. Local grazing practices appear to have had a role in both the spread and control of the outbreak. Goats are an important form of savings and cash income to people in the locality, and the outbreak may result in considerable financial hardship for affected goat keepers. We provide a detailed description of the clinical disease and the spread of the outbreak in the locality. Awareness of the disease with reference to farming practices will provide opportunities for future disease control to enhance animal welfare and rural prosperity.

The role of goats as reservoir hosts for bovine herpes virus 1 under field conditions.

Bovine herpesvirus 1 (BoHV1) is the cause of economically significant viral infections in cattle. Respiratory symptoms associated with the infection are known as Infectious Bovine Rhinotracheitis (IBR). Sheep and goats are less sensitive to the infection although their role in inter-species viral transmission under field conditions is subject to controversy. The objective of this study was to investigate seroprevalence of BoHV1 infections in cattle, sheep, and goats raised together for at least a year. Blood serum samples were taken from 226 cattle, 1.053 sheep, and 277 goats from 17 small- to medium-scale farms. BoHV1-specific antibody presence and titers were determined using virus neutralization test. In total, 73 of the 226 cattle (32.3%) were seropositive. The infection was detected in 13 of the 17 farms. Infection rates ranged from 5.8 to 88.8%. Only one of the 1053 sheep (0.09%) was seropositive. However, 58 of the 277 (20.9%) goats were seropositive. Goat samples taken from 8 of the 17 farms were seropositive with infection rates ranging from 17 to 38.9%. Statistical analysis showed a significant correlation in infection rates between cattle and goats but not sheep. These results suggest that goats may be more sensitive to the BHV1 infection than sheep and the role of goats as possible reservoirs for BoHV1 in the control and eradication of BHV1 in cattle should be considered in future studies.

Improving Quality Control of Contagious Caprine Pleuropneumonia Vaccine with Tandem Mass Spectrometry.

Vaccines to protect livestock against contagious caprine pleuropneumonia (CCPP) consist of inactivated, adjuvanted antigens. Quality control of these vaccines is challenging as total protein quantification provides no indication of protein identity or purity, and culture is not an option. Here, a tandem mass spectrometry approach is used to identify the mycoplasma antigen contained in reference samples and in commercial CCPP vaccines. By the same approach, the relative amounts of mycoplasma antigen and residual proteins originating from the production medium are determined. Mass spectrometry allows easy and rapid identification of the peptides present in the vaccine samples. Alongside the most probable mycoplasma species effectively present in the vaccines, a very high proportion of peptides from medium constituents are detected in the commercial vaccines tested.

Identification of Peste des Petits Ruminants Virus, Georgia, 2016.

A phylogenetic analysis of samples taken from reported outbreaks of peste des petits ruminants virus (PPRV) in Georgia revealed a closer relationship to viruses from northern and eastern Africa than to viruses from countries closer to Georgia. This finding has crucial implications for the control of PPRV in the region.

Complete genome characterization of a rotavirus B (RVB) strain identified in Alpine goat kids with enteritis reveals inter-species transmission with RVB bovine strains.

Rotavirus B (RVB) has been associated with enteric disease in many animal species. An RVB strain was identified in pooled intestinal samples from Alpine caprine kids (between 2 and 3 days of age) experiencing high (>90 %) morbidity, and the complete caprine RVB genome was characterized. Histology revealed villus atrophy, the samples tested positive for RVB by real-time RT-PCR and metagenomic next-generation sequencing identified only RVB and orf virus. In the VP4 gene segment, the caprine RVB strain had a higher percentage nucleotide identity to the Indian bovine RVB strains than to the Japanese bovine RVB strains, but the VP7, VP6, VP2, NSP1, NSP2 and NSP5 gene segments of the American caprine RVB strain were genetically related to the Japanese bovine RVB strains. The results indicate a lack of RVB sequences to understand reassortment or the evolutionary relationship of RVB strains from cattle and goats.

Sheep as a Potential Source of Zoonotic Cryptosporidiosis in China.

In this study, we assessed the prevalence and genetic characteristics of Cryptosporidium in sheep from 10 provinces in China. Fecal samples from 1,035 sheep originating from 16 farms were collected, and 295 (28.5%) were found to be Cryptosporidium positive by nested PCR. Cryptosporidium was detected at all farms, with infection rates between 5.7% and 50.0%. Three Cryptosporidium species were identified, including Cryptosporidium xiaoi (73.2%, 216/295), Cryptosporidium ubiquitum (21.7%, 64/295), and Cryptosporidium parvum (5.1%, 15/295). The distribution of Cryptosporidium species differed by province and by farm. All three species were detected in lambs and adult sheep but the highest infection rate was found in postweaned lambs. All three species were detected in all four seasons, with the highest prevalence found in autumn. Four C. parvum subtypes (IIaA15G2R1, IIaA17G2R1, IIdA18G1, and IIdA19G1) and one C. ubiquitum subtype (XIIa) were identified. For most provinces in this study, we are not aware of a previously published description or molecular characterization of Cryptosporidium infections in sheep. This information will improve our knowledge and understanding of the epidemiology of cryptosporidiosis in China.IMPORTANCECryptosporidium is an important zoonotic parasite that causes diarrhea in humans and animals worldwide. Previous studies suggested geographic differences in the distribution of Cryptosporidium species in sheep. However, molecular characterization studies of Cryptosporidium species in sheep have been carried out in only a few provinces in China, and the limited data available do not reflect the real situation. In this study, five districts, covering most areas where sheep are bred in China, were selected for examination of Cryptosporidium species, and Cryptosporidium infections were detected at all farms assessed, suggesting that Cryptosporidium is widespread in sheep in China. We also found geographic differences in the distribution of Cryptosporidium species but did not detect any differences between sheep age groups or seasons. Subtyping analyses showed that all of the subtypes identified in this study have been reported in humans, suggesting that sheep may be a potential source of zoonotic cryptosporidiosis.

Molecular verification of transplacental transmission of Theileria lestoquardi in goat.

Ovine and caprine malignant theileriosis (OCMT), a critical condition in small ruminant production, causes lethal infections. In September 2016, a total number of 400 goats of Marghoz breed (the largest population of goat breed in Iran), in northwest of Iran, were examined for approximately 25 pre-partum abortions and 7 goats' mortality for a period of 3 days. A dead goat and her aborted fetus were brought into the Urmia Veterinary Hospital for further diagnostic investigations. The microbiological assessments including direct microscopical examination of the vaginal discharges and placentome with respect to the differential staining (Gram's staining method), conventional pure culturing, fetal abomasal contents, and the liver were negative. Microscopic examinations of blood smears of the goat and the fetus revealed characteristic of Theileria spp. piroplasms and impression smear samples from goat liver and fetal spleen were positive for Theileria Koch blue bodies. DNA analysis was performed using PCR technique and specific primers derived from the nucleotide sequences of 18S rRNA gene of T. lestoquardi; following extraction from blood samples, placentome, goat liver, and spleen of the fetus. The amplified DNA was sequenced afterwards and the corresponding sequence was registered under GenBank accession number MG208059. The sequence alignment showed that the products of PCR had a homology of 99% to known T. lestoquardi sequence registered under accession numbers of KY352037.1, KC778786.1, and JQ917458.1 in the GenBank. To our knowledge, this is a report demonstrating molecular verification of T. lestoquardi transplacental transmission in a neonatal kid of Marghoz breed of goats, its feasible role in induction of perinatal deaths and abortion in goat flocks.

Rodents, goats and dogs - their potential roles in the transmission of schistosomiasis in China.

Schistosomiasis in China has been substantially reduced due to an effective control programme employing various measures including bovine and human chemotherapy, and the removal of bovines from endemic areas. To fulfil elimination targets, it will be necessary to identify other possible reservoir hosts for Schistosoma japonicum and include them in future control efforts. This study determined the infection prevalence of S. japonicum in rodents (0-9·21%), dogs (0-18·37%) and goats (6·9-46·4%) from the Dongting Lake area of Hunan province, using a combination of traditional coproparasitological techniques (miracidial hatching technique and Kato-Katz thick smear technique) and molecular methods [quantitative real-time PCR (qPCR) and droplet digital PCR (ddPCR)]. We found a much higher prevalence in goats than previously recorded in this setting. Cattle and water buffalo were also examined using the same procedures and all were found to be infected, emphasising the occurrence of active transmission. qPCR and ddPCR were much more sensitive than the coproparasitological procedures with both KK and MHT considerably underestimating the true prevalence in all animals surveyed. The high level of S. japonicum prevalence in goats indicates that they are likely important reservoirs in schistosomiasis transmission, necessitating their inclusion as targets of control, if the goal of elimination is to be achieved in China.

Codon 141 polymorphisms of the ovine prion protein gene affect the phenotype of classical scrapie transmitted from goats to sheep.

BACKGROUND: A study to investigate transmission of classical scrapie via goat milk was carried out in sheep: firstly, lambs were challenged orally with goat scrapie brain homogenate to confirm transmission of scrapie from goats to sheep. In the second study phase, milk from scrapie-infected goats was fed to lambs. Lambs were selected according to their prion protein gene (PRNP) genotype, which was either VRQ/VRQ or ARQ/ARQ, with or without additional polymorphisms at codon 141 (FF141, LF141 or LL141) of the ovine PRNP. This report describes the clinical, pathological and molecular phenotype of goat scrapie in those sheep that progressed to clinical end-stage. RESULTS: Ten sheep (six VRQ/VRQ and four ARQ/ARQ, of which three FF141 and one LL141) challenged with one of two scrapie brain homogenates, and six pairs of sheep (ARQ, of which five LL141 and seven LF141) fed milk from six different goats, developed clinical disease, which was characterised by a pruritic (all VRQ/VRQ and LL141 sheep) or a non-pruritic form (all LF141 and FF141 sheep). Immunohistochemical (IHC) examination revealed that the pattern of intra- and extracellular accumulation of disease-associated prion protein in the brain was also dependent on PRNP polymorphisms at codon 141, which was similar in VRQ and LL141 sheep but different from LF141 and FF141 sheep. The influence of codon 141 was also seen in discriminatory Western blot (WB), with LF141 and FF141 sheep showing a bovine spongiform encephalopathy-like profile (diminished reactivity with P4 antibody) on brain tissue. However, discriminatory WB in lymphoid tissues, and IHC pattern and profile both in lymphoid and brain tissue was consistent with classical scrapie in all sheep. CONCLUSIONS: This study provided further evidence that the clinical presentation and the pathological and molecular phenotypes of scrapie in sheep are influenced by PRNP polymorphisms, particularly at codon 141. Differences in the truncation of disease-associated prion protein between LL141 sheep and those carrying the F141 allele may be responsible for these observations.

Infection levels of protostrongylid nematodes in definitive caprine and intermediate gastropod hosts from Uzbekistan.

Morphological analysis of lungworms collected among Caprinae from Uzbekistan resulted in the identification of four species of Protostrongylidae: Protostrongylus rufescens, Protostrongylus hobmaieri, Spiculocaulus leuckarti and Cystocaulus ocreatus. The following species were recorded as definitive hosts: Ovis aries, Ovis ammon, Ovis vignei, Capra hircus, Capra falconeri and Capra sibirica. The prevalence of P. rufescens reached 45.3%, followed by S. leuckarti and C. ocreatus with 31.7% and P. hobmaieri with 16.9%. The sex ratio ranged between 1:3.1 and 1:6.2, with P. hobmaieri showing the strongest predominance of females over males. The prevalence of infection of small ruminants with protostrongylid nematodes increased with the age of the hosts. Protostrongyles use terrestrial gastropods as intermediate hosts, and infective larvae were found in the species Vallonia costata, Gibbulinopsis signata, Pupilla muscorum, Pseudonapaeus albiplicata, Pseudonapaeus sogdiana, Leucozonella ferghanica, Xeropicta candacharica, Candaharia levanderi and Macrochlamys sogdiana. Xeropicta candacharica was the most abundant gastropod and had the highest prevalence of infection with protostrongylids. Adult X. candacharica had a significantly higher infection intensity than juveniles. The epidemiology of protostrongylid infections is dynamic and subject to considerable changes. Further characterization of the interaction of protostrongylid parasites with their terrestrial gastropods as intermediate hosts and Caprinae as definitive hosts is required to understand these processes and to monitor the effects of changing ecological contexts.

Genetic and Pathological Follow-Up Study of Goats Experimentally and Naturally Exposed to a Sheep Scrapie Isolate.

UNLABELLED: Thirty-seven goats carrying different prion protein genotypes (PRNP) were orally infected with a classical scrapie brain homogenate from wild-type (ARQ/ARQ) sheep and then mated to obtain 2 additional generations of offspring, which were kept in the same environment and allowed to be naturally exposed to scrapie. Occurrence of clinical or subclinical scrapie was observed in the experimentally infected goats (F0) and in only one (F1b) of the naturally exposed offspring groups. In both groups (F0 and F1b), goats carrying the R154H, H154H, R211Q, and P168Q-P240P dimorphisms died of scrapie after a longer incubation period than wild-type, G37V, Q168Q-P240P, and S240P goats. In contrast, D145D and Q222K goats were resistant to infection. The immunobiochemical signature of the scrapie isolate and its pathological aspects observed in the sheep donors were substantially maintained over 2 goat generations, i.e., after experimental and natural transmission. This demonstrates that the prion protein gene sequence, which is shared by sheep and goats, is more powerful than any possible but unknown species-related factors in determining scrapie phenotypes. With regard to genetics, our study confirms that the K222 mutation protects goats even against ovine scrapie isolates, and for the first time, a possible association of D145 mutation with scrapie resistance is shown. In addition, it is possible that the sole diverse frequencies of these genetic variants might, at least in part, shape the prevalence of scrapie among naturally exposed progenies in affected herds. IMPORTANCE: This study was aimed at investigating the genetic and pathological features characterizing sheep-to-goat transmission of scrapie. We show that in goats with different prion protein gene mutations, the K222 genetic variant is associated with scrapie resistance after natural and experimental exposure to ovine prion infectivity. In addition, we observed for the first time a protective effect of the D145 goat variant against scrapie. Importantly, our results demonstrate that the phenotypic characteristic of the wild-type sheep scrapie isolate is substantially preserved in goats carrying different susceptible PRNP gene variants, thus indicating that the prion protein gene sequence, which is shared by sheep and goats, plays a fundamental role in determining scrapie phenotypes.

Experimental caprine neosporosis: the influence of gestational stage on the outcome of infection.

Here, we assessed outcome of experimental infection by Neospora caninum in goats intravenously inoculated with 10(6) tachyzoites of the Nc-Spain7 isolate at 40 (G1), 90 (G2) and 120 (G3) days of gestation. Infected goats had fever between 5 and 9 days post inoculation (dpi); all were seropositive at the time of abortion/birth. Foetal death occurred in G1 from 10 to 21 dpi (n = 7) and in G2 from 27 to 35 dpi (n = 4). Goats in G2 also had seropositive stillbirth (n = 1) and healthy kids (n = 2). G3 goats (n = 7) had 3 seropositive and 3 seronegative weak kids, and 2 seronegative healthy kids. Parasite DNA detection in placentomes was 100% in G2, 85.7% in G3 and in G1 was detected only in placentomes from the goats with foetal losses from 17 dpi (100%). Parasites were detected in foetal/kid brain (>85.7%) and liver (≥ 50%) of G2 and G3, and in G1 after 17 dpi (100%). The highest parasite loads were detected in the placentomes of G1 from 17 dpi and G2, and in foetal tissues of G1 from 17 dpi and G3. Multifocal necrotic lesions were observed in the placentas of the three groups, but they were larger and more frequent in G1 and G2. Similar lesions were observed in foetal tissues, but they were more frequent in G3. These findings suggest that, as observed in cattle and sheep, the clinical consequences of N. caninum in pregnant goats are dependent in part on the time of gestation when animals were infected.

Integrating serological and genetic data to quantify cross-species transmission: brucellosis as a case study.

Epidemiological data are often fragmented, partial, and/or ambiguous and unable to yield the desired level of understanding of infectious disease dynamics to adequately inform control measures. Here, we show how the information contained in widely available serology data can be enhanced by integration with less common type-specific data, to improve the understanding of the transmission dynamics of complex multi-species pathogens and host communities. Using brucellosis in northern Tanzania as a case study, we developed a latent process model based on serology data obtained from the field, to reconstruct Brucella transmission dynamics. We were able to identify sheep and goats as a more likely source of human and animal infection than cattle; however, the highly cross-reactive nature of Brucella spp. meant that it was not possible to determine which Brucella species (B. abortus or B. melitensis) is responsible for human infection. We extended our model to integrate simulated serology and typing data, and show that although serology alone can identify the host source of human infection under certain restrictive conditions, the integration of even small amounts (5%) of typing data can improve understanding of complex epidemiological dynamics. We show that data integration will often be essential when more than one pathogen is present and when the distinction between exposed and infectious individuals is not clear from serology data. With increasing epidemiological complexity, serology data become less informative. However, we show how this weakness can be mitigated by integrating such data with typing data, thereby enhancing the inference from these data and improving understanding of the underlying dynamics.

Outbreak of Occupational Brucellosis in a Laboratory Technician at Her Royal Highness Princess Sirindhorn Medical Center, Srinakharinwirot University, Thailand.

Objective: To document laboratory transmission of brucellosis and identify the likely mechanism of transmission of brucellosis at Her Royal Highness (HRH) Princess Sirindhorn Medical Center, Thailand. Material and Method: Using small subunit ribosomal RNA (rRNA) sequencing technique to analyze Brucella melitensis cultured from the first 2 patients of the hospital and an infected laboratory technician, and using brucellosis serologic test to rule out infections in all other involved technicians. Results: We had encountered the first 2 cases of brucellosis. Both had infected from community exposure with goat. The first case had pancreatic abscess and spinal bone involvement with a positive blood culture. The second case presented with fever of unknown origin and had a positive blood culture. A few weeks later, 1 of our laboratory technicians presented with fever, myalgia and fatigue. Blood culture grew B. melitensis. He never had any associated community-acquired risk factors for brucellosis. The presumed mechanism of transmission was an inhalation while taking photographs of the bacterial plate of the first patient. B. melitensis identified from our laboratory technician and both patients were analyzed based on 16S-23S rRNA intergenic transcribed spacer (ITS) region. Results of 16S-23S rRNA ITS sequence testing confirmed a match from all patients and laboratory technician's isolate. All other 10 potentially exposed laboratory technicians were asymptomatic. A brucellosis serologic test was negative in all non-infected technicians but was only positive in the 1 infected technician. Conclusion: This is the first report in Thailand of occupational brucellosis transmitted in microbiologic laboratory. The most likely mechanism is air-borne inhalation of bacterial organisms on culture media in the absence of adequate precautions. Laboratory technicians should handle Brucella cultivation with caution utilizing appropriate measures to prevent inhalation.