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  1. Abdul Rahman MS, Khor KH, Khairani-Bejo S, Lau SF, Mazlan M, Roslan MA
    Animals (Basel), 2021 Nov 29;11(12).
    PMID: 34944181 DOI: 10.3390/ani11123405
    Canine leptospirosis is commonly associated with kidney and/or liver disease. It has been widely reported and causes public health concerns due to its zoonotic potential and its re-emergence, resulting from close contact between humans and dogs. The current study identified potential risk and predictive factors for dogs diagnosed with kidney and/or liver disease due to leptospirosis. A total of 124 client-owned dogs were recruited, and information such as signalment, medical history, management, and clinical findings were documented. Samples collected from the recruited dogs were directly tested using polymerase chain reaction (PCR) and subsequently inoculated for bacterial isolation. Statistical analyses were descriptively analyzed, and risk analyses were performed using Pearson chi-square tests and logistic regression. A total of 53 dogs (42.7%) were positive for leptospiral infection based on PCR, and 10 leptospiral isolates were successfully recovered from eight dogs. The mortality rate of infected dogs was 34.0% (18/53). Medium and large dog breeds, with a history of exposure to rats, and managed outdoors had a greater risk for leptospirosis (p < 0.05). The significant predictors for the dogs' positivity were the presence of rats and acute clinical illness (p < 0.05). Administration of antibiotics and the detection of clinical illness at an early stage of the disease improved the survivability of the dogs (p < 0.05). Identifying the profile of dogs that are at risk to leptospirosis could be useful in the design of diagnostic and treatment strategies, as well as to increase awareness for prevention of the disease.
  2. Ahmed IM, Khairani-Bejo S, Hassan L, Bahaman AR, Omar AR
    BMC Vet Res, 2015;11:275.
    PMID: 26530141 DOI: 10.1186/s12917-015-0587-2
    Brucella melitensis is the most important pathogenic species of Brucella spp. which affects goats and sheep and causes caprine and ovine brucellosis, respectively. Serological tests for diagnosis of brucellosis such as Rose Bengal plate test (RBPT) and enzyme-linked immunosorbent assay (ELISA) usually utilize smooth lipopolysaccharides (S-LPS) as a diagnostic antigen which could give false positive serological reactions. Outer membrane proteins (OMP) of B. melitensis have been used as alternative diagnostic antigens rather than S-LPS for differential serological diagnosis of brucellosis, mainly in ELISA with single recombinant OMP (rOMP) as a diagnostic antigen. Nevertheless, the use of single format mainly showed lack of sensitivity against the desired rOMP. Therefore, this study aimed to determine whether a newly developed rOMPs indirect ELISA (rOMPs I-ELISA), based on combination of rOMP25, rOMP28 and rOMP31of B. melitensis, has a potential benefit for use in the serodiagnosis of brucellosis.
  3. Alashraf AR, Lau SF, Khor KH, Khairani-Bejo S, Bahaman AR, Roslan MA, et al.
    Top Companion Anim Med, 2019 Mar;34:10-13.
    PMID: 30808490 DOI: 10.1053/j.tcam.2018.12.002
    Leptospirosis is one of the most widespread zoonotic diseases and despite extensive research, there is still a paucity of information regarding this disease in cats. The aim of this study was to investigate the prevalence of leptospirosis among the shelter cat population in Malaysia and to determine the most common infective Leptospira serogroups among them. Blood samples were collected from a total of 110 cats from 4 different shelters. The sampled cats appeared healthy, with minimal evidence of feline upper respiratory disease. The Microscopic Agglutination Test was used to detect anti-Leptospira antibodies against 20 pathogenic serovars. Based on a cut-off antibody titer of ≥1:100, 20 of 110 sheltered cats, showed presence of anti-Leptospira antibodies against at least 1 serovar. The serodetection of leptospirosis was 18.18% (95% confidence interval 12.09-26.42). The most commonly detected serogroups were Bataviae, Javanica, and Ballum, with antibody titers ranging from 1:100 to 1:1600. Knowledge of the predominant infective serovars in hosts worldwide and regionally is imperative for understanding the epidemiology of this zoonotic disease. Serosurveillance is the first step in this process. Further studies are warranted for investigation of urinary shedding in naturally infected cats with leptospirosis, using Polymerase chain reaction (PCR) and organism isolation followed by serovars identification.
  4. Alashraf AR, Lau SF, Khairani-Bejo S, Khor KH, Ajat M, Radzi R, et al.
    PLoS One, 2020;15(3):e0230048.
    PMID: 32155209 DOI: 10.1371/journal.pone.0230048
    Leptospirosis is one of the most widespread zoonotic diseases and can infect both humans and animals worldwide. Healthy cat, as a potential source of exposure to humans, are likely underestimated owing to the lack of overt clinical signs associated with Leptospira spp. infection in this species. The aim of the study was to determine the exposure, shedding, and carrier status of leptospires in shelter cats in Malaysia by using serological, molecular, and bacteriological methods. For this study, 82 healthy cats from two shelters were sampled. The blood, urine, and kidneys were tested using the microscopic agglutination test (MAT), polymerase chain reaction (PCR), and bacterial culture. On the basis of serological, molecular, and/or culture techniques, the total detection of leptospiral infection was 29.3% (n = 24/82). Through culture techniques, 16.7% (n = 4/24) of the cats that tested positive were carriers with positive kidney cultures, and one cat was culture positive for both urine and kidney. The Leptospira spp. isolates were identified as pathogenic L. interrogans serovar Bataviae through serological and molecular methods. Through serological techniques, 87.5% (n = 21/24) had positive antibody titers (100-1600) and most of the Bataviae serogroup (n = 19/21). Using PCR, 16.7% (n = 4/24) of cats were shown to have pathogenic Leptospira spp. DNA in their urine. Furthermore, three out of four culture positive cats were serology negative. The present study reports the first retrieval of pathogenic leptospires from urine and kidneys obtained from naturally infected cats. The results provide evidence of the potential role of naturally infected cats in the transmission of leptospires. Additionally, leptospiral infection occurs sub-clinically in cats. The culture isolation provides evidence that healthy cats could be reservoirs of leptospiral infection, and this information may promote the development of disease prevention strategies for the cat population.
  5. Anka MS, Hassan L, Khairani-Bejo S, Zainal MA, Mohamad RB, Salleh A, et al.
    PLoS One, 2014;9(9):e108673.
    PMID: 25265020 DOI: 10.1371/journal.pone.0108673
    Bovine brucellosis was first reported in Peninsular Malaysia in 1950. A subsequent survey conducted in the country revealed that the disease was widespread. Current knowledge on the potential risk factors for brucellosis occurrence on cattle farms in Malaysia is lacking. Therefore, we conducted a case-control study to identify the potential herd-level risk factors for bovine brucellosis occurrence in four states in the country, namely Kelantan, Pahang, Selangor and Negeri Sembilan. Thirty-five cases and 36 controls of herds were selected where data on farm management, biosecurity, medical history and public health were collected. Multivariable logistic regression identified that Brucella seropositive herds were more likely to; have some interaction with wildlife (OR 8.9, 95% CI = 1.59-50.05); originated from farms where multiple species such as buffalo/others (OR 41.8, 95% CI = 3.94-443.19) and goat/sheep (OR 8.9, 95%Cl = 1.10-71.83) were reared, practice extensive production system (OR 13.6, 95% CI 1.31-140.24) and have had episodes of abortion in the past (OR 51.8, 95% CI = 4.54-590.90) when compared to seronegative herds. Considering the lack of information on the epidemiology of bovine brucellosis in peninsular Malaysia and absence of information on preventing the inception or spread of the disease, this report could contribute to the on-going area-wise national brucellosis eradication program.
  6. Anka MS, Hassan L, Adzhar A, Khairani-Bejo S, Mohamad RB, Zainal MA
    BMC Vet Res, 2013;9:230.
    PMID: 24246015 DOI: 10.1186/1746-6148-9-230
    Bovine brucellosis is an important disease affecting cattle characterised by abortion, still birth, reduced milk production, weak foetus and infertility in both males and females. There is wide distribution of the disease among cattle and several wildlife species. Bovine brucellosis is commonly caused by B. abortus and very occasionally B. melitensis and B. suis. The distribution of bovine brucellosis in cattle has not been described in Malaysia. In this paper we describe the distribution, pattern and trend of bovine brucellosis in Peninsular Malaysia between 2000 and 2008 based on serological data obtained from nationwide B. abortus serosurveillance activities in cattle populations.
  7. Bahaman, A.R., Kattan, G, Khairani- Bejo, S., Mutalib, A.R., Zakaria, Z.
    Jurnal Veterinar Malaysia, 2017;29(2):1-4.
    MyJurnal
    Leptospirosis is re-emerging in Malaysia and many other tropical countries. Its infection of human and animal’s worldwide. The study was carried out to identify predominate Leptospira serovars within rat populations in Kuala Lumpur (KL) by microscopic agglutination test (MAT). As well to investigate the frequency of infection by polymerase chain reaction assays (PCR). The isolated cultures that obtained from culturing kidney tissue of 112 trapped rats from four sites in KL, were identified by MAT using panel of 16 standard hyper immune anti-sera. Besides, identification of Leptospira strains by PCR amplification with G1/G2 and ompL1 Borgpetersenii genes. Results from typing by MAT revealed that 8/57 (7.1%) of cultured isolates reacted against; hyper-immune antisera of serogroup Javanica and 13/57 (22.8%) against serogroup Bataviae. Whereas, the rest of isolates recorded across reactivity 1/20 against serogroups; Icterohaemorrhagiae 2/57, Canicola 1/57, Australis 2/57. From PCR-G1/G2; 50/112 (50.9%) cultures were positive included (40/57 cultures positive to isolation and 10/55 cultures negative to isolation). While, from PCR-ompL1 Borgpetersenii gene; 21/112 (18.7%) cultures positive included (17/57 cultures positive to isolation and 4/55 cultures were negative to isolation). It is obvious from this study that serogroups Javanica and Bataviae were the predominant among rat populations in KL. Also it revealed the high frequency of pathogenic strains among rat populations and their potential risk of humans and animals contracting infection.
  8. Bamaiyi PH, Hassan L, Khairani-Bejo S, ZainalAbidin M, Ramlan M, Adzhar A, et al.
    Prev Vet Med, 2015 May 1;119(3-4):232-6.
    PMID: 25746928 DOI: 10.1016/j.prevetmed.2015.02.001
    A study was conducted to describe the prevalence and distribution of zoonotic Brucella melitensis in goats in Peninsular Malaysia. Using serosurveillance data of the last decade (2000-2009) involving 119,799 goats and 3555 farms, the seroprevalence of brucellosis among goats was 0.91% (95% CI=0.86-0.96) and among farms was 7.09% (95% CI=6.27-7.98). The odds of brucellosis was significantly (P<0.05) higher in the later part of the decade, in larger herd size and among the states located in the peninsula as compared to eastern Malaysia. The infection was detected throughout Malaysia but at generally low seroprevalences with states like Perlis that border neighbouring countries having higher seroprevalence of brucellosis than other non-border states.
  9. Bamaiyi PH, Hassan L, Khairani-Bejo S, ZainalAbidin M, Ramlan M, Krishnan N, et al.
    Trop Anim Health Prod, 2014 Jun;46(5):739-45.
    PMID: 24633657 DOI: 10.1007/s11250-014-0557-x
    Caprine brucellosis is a bacterial zoonotic infection affecting goats especially in developing countries all over the world. In Malaysia, the risk factors associated with this infection in farms have not been studied. A case-control study was carried out in goat farms in four states of Malaysia to elucidate the risk factors associated with the infection on the farms using structured questionnaires and face-to-face interviews. Results indicate that the introduction of new animals (OR = 5.25; 90 % CI = 1.46, 18.88), younger age category of farms (OR = 5.53; 90 % CI = 1.09, 21.66), and farms with single breed of goats (OR = 8.50; 90 % CI = 1.27, 41.97) were significant risk factors for brucellosis. In order to control brucellosis or possibly eradicate it in goat farms, these factors need to be dealt with. Enforcing stringent importation protocols or complete ban of goat importation from brucellosis endemic countries will help reduce risk of introducing new infection into the country.
  10. Bamaiyi PH, Hassan L, Khairani-Bejo S, Zainal Abidin M, Ramlan M, Krishnan N, et al.
    Trop Biomed, 2012 Dec;29(4):513-8.
    PMID: 23202595
    A study was carried out to isolate Brucella melitensis using established bacteriological and PCR techniques in Brucella seropositive goats in farms in Selangor, Negeri Sembilan, Melaka and Pulau Pinang. Brucella melitensis was isolated from 7 of 134 reactors with the highest isolation from the vaginal swabs (57.14%) followed by the spleen (28.57%), uterine fluid (14.29%). No Brucella was isolated from the lymph nodes. PCR confirmed all the seven isolates as B. melitensis and isolates were phylogenetically related to other isolates from India, Iran, and Israel but most closely related to isolates from Singapore.
  11. Bitrus AA, Zunita Z, Khairani-Bejo S, Othman S, Ahmad Nadzir NA
    Microb Pathog, 2018 Oct;123:323-329.
    PMID: 30053600 DOI: 10.1016/j.micpath.2018.07.033
    This study was designed to screen for SCCmec types and to characterize the attachment site (attB) and universal insertion site (orfX) of SCCmec in a collection of 27 isolates (n = 11) methicillin resistant S. aureus and (n = 16) methicillin susceptible S. aureus isolates in Malaysia. Screening of SCCmec types and characterization of the attachment site was carried out using PCR amplification and Sanger's sequencing method. The result showed that a large proportion of the MRSA isolates carried SCCmec type III 7/11 (63%). Three isolates 3/11 (27%) and 1/11 (9.0%) carried SCCmec type II and IVd respectively. Amplification of the universal insertion site of the SCCmec (orfX) and attachment site (attB) showed that all 16 S. aureus isolates were positive for the orfX gene, while only 7 were positive for the attB gene. Phylogenetic diversity showed that the isolates clustered around strains with features similar to a community acquired MRSA. In conclusion, a high carriage rate of SCCmec type III was observed. The result also showed that all the S. aureus isolates have the orfX structure; however, not all isolates possesses the attB site on the 3' end of the orfX region.
  12. Garba B, Bahaman AR, Khairani-Bejo S, Zakaria Z, Mutalib AR
    Ecohealth, 2017 Jun;14(2):389-398.
    PMID: 28405850 DOI: 10.1007/s10393-017-1234-0
    Leptospirosis is a bacterial disease transmitted to humans and animals by direct or indirect contact with urine or body fluids from infected animals especially rodents. Infection can be associated with wide clinical spectrum varying from asymptomatic to severe multi-organ syndrome with life-threatening consequences. We conducted a review of published studies on incidences, case reports, sero-epidemiological surveys from year 2000 to 2015 using different electronic data bases. Our study revealed that majority of the studies were conducted in Peninsular Malaysia and predominantly among high-risk human groups. Most of the studies on domestic animals were conducted in the 1980s; hence, the current status of leptospirosis among domestic animal population remains largely unknown. There tend to be a sharp rise in incidence rate among human population in the year 2014 which was attributed to flooding and heavy rainfall experienced as well as recreational activities. Several gaps in epidemiological knowledge were also disclosed.
  13. Goh SH, Khor KH, Ismail R, Megat Abdul Rani PA, Mohd Mohidin TB, Bahaman AR, et al.
    Trop Biomed, 2020 Dec 01;37(4):1074-1082.
    PMID: 33612759 DOI: 10.47665/tb.37.4.1074
    The incidence of leptospirosis seems to be on the rise and could be an alarming indirect indication of a global re-emergence. It is a potential public health threat when dogs are speculated to be involved in the transmission of leptospirosis through possible subclinical harbouring of Leptospira spp. and subsequent shedding into the environment. This study aimed to detect anti-leptospiral antibodies among dogs and their handlers using the microscopic agglutination test (MAT). Blood samples from 266 apparently healthy dogs and 194 dog handlers were collected at four working dog organisations and four dog shelters. Serum samples were tested using MAT against 20 leptospiral serovars with a cut-off titre >=1:100 (dog) and >=1:50 (dog handlers). Seventy dogs (70/266; 26.3%) were seropositive mainly against serovars Icterohaemorrhagiae, Ballum, Bataviae and Javanica (titres ranged: 1:100-1:800). Sixty-seven dog handlers (67/194; 34.5%) were seropositive mainly against serovars Grippotyphosa, Icterohaemorrhagiae and Malaysia (titres ranged: 1:50-1:200). Dogs were seropositive due to exposure, vaccination or active infection. Seropositive dog handlers could indicate exposure or active infection. This shows the potential of dogs in maintaining and spreading the infection in Malaysia. Due to the occupational risk as a result of frequent contact with dogs and exposure to contaminated environments, dog handlers should be made aware of the presence of this zoonotic disease.
  14. Goh SH, Khor KH, Radzi R, Lau SF, Khairani-Bejo S, Abdul Rahman MS, et al.
    Top Companion Anim Med, 2021 Nov;45:100562.
    PMID: 34271178 DOI: 10.1016/j.tcam.2021.100562
    Leptospirosis is an endemic zoonoses of global proportions. Stray dogs have been postulated to play a role in disease transmission; however, supporting information are still limited. Roaming behavior may not only predispose the dogs to infection, but could also contribute to disease spread. In this study, the susceptibility of urban stray dogs in shedding Leptospira spp. was determined. Blood, urine, and tissue samples of kidney and liver were collected from 100 dogs from 2 animal control facilities. Serological testing using microscopic agglutination test (MAT) were performed on blood against 20 leptospiral serovars with a cut-off titre of ≥ 1:100. Samples were cultured onto semi-solid Ellinghausen and McCullough modified by Johnson and Harris (EMJH) media. Isolates were identified using molecular polymerase chain reaction (PCR) using 2 primers (16s rRNA and LipL32) and hyperimmune serum (HIS) MAT. The seroprevalence for the dogs positive for leptospirosis was 32% (n=32/100) with the following detected serovars: Javanica (n=13), Bataviae (n=10), Icterohaemorrhagiae (n=3), Autumnalis (n=2), Canicola (n=1), Pyrogenes (n=1), Copenhageni (n=1), and Australis (n=1). Six Leptospira spp. isolated were procured from urine (n=2), kidney (n=2) and liver (n=2). All 6 isolates belonged to L. interrogans, a pathogenic variant of Leptospira spp. Serotyping and phylogenetic analysis suggested serovar Bataviae (n=5) and serovar Canicola (n=1). Presence of vaccinal serovars (Icterohaemorrhagiae and Canicola) suggested potential post-vaccination antibodies but the predominance of non-vaccinal serovars (Javanica and Bataviae) indicate the possibility of current infection or post-exposure. Isolation of Leptospira spp. directly from urine sample not only suggested an active infection but highlighted the potential shedding capability among these stray dogs. These findings further strengthen speculations that urban stray dogs could play a role in transmission and dissemination of leptospirosis through their constant movement. The urine of infected dogs may contaminate the environment, posing a major public health threat.
  15. Goh SH, Ismail R, Lau SF, Megat Abdul Rani PA, Mohd Mohidin TB, Daud F, et al.
    PMID: 31035316 DOI: 10.3390/ijerph16091499
    This study determined the potential risk factors that may contribute to seropositivity among dogs and dog handlers from working dog and dog shelter institutions. Data was collected from dogs (n = 266) and dog handlers (n = 161) using a standardised guided questionnaire. Serum obtained from the dogs and dog handlers was tested using the microscopic agglutination test (MAT). A logistic regression analysis was used to predict leptospiral seropositivity of dogs and dog handlers based on potential risk factors. A total of 22.2% of dogs and 21.7% of dog handlers were seropositive. The significant predictors for the dogs' seropositivity were presence of rats (OR = 4.61 (95% CI: 1.05, 20.33), p = 0.043) and shared common area (OR = 5.12 (95% CI: 1.94, 13.46), p = 0.001) within the organisation. Significant predictor for dog handler seropositivity was contact time with the dogs of more than six hours/day (OR = 3.28 (95% CI: 1.28, 8.40), p = 0.013) after controlling for the effect of other risk factors such as small mammal contact, rat infestation at home, flooding at housing area (within three months) and urban locality. The exposure to various disease sources identified poses risk to dogs and dog handlers. Risk could be reduced with adequate application of protection at work while handling dogs and thus limiting contact with these sources and reducing exposure to infection.
  16. Goh, S.H., Khor, K.H., Lau, S.F., Ismail, R., Khairani-Bejo, S., Radzi, R.
    Jurnal Veterinar Malaysia, 2019;31(1):17-27.
    MyJurnal
    Dogs could be reservoirs and carriers of Leptospira spp., hence, may play a potential role in disease transmission, exposing dog handlers to leptospirosis. This study evaluated the dog handlers’ knowledge, attitude and practice (KAP) towards leptospirosis. Four working dog and four dog shelter organisations were approached. A total of 138 dog handlers were conveniently recruited and evaluated on their level of knowledge, attitude and practice towards leptospirosis using a validated structured questionnaire. Most of the dog handlers were male aged between 30 and 40 years with a wide range of working experience of 1 - 10 years and all of them have no formal training about leptospirosis. The majority of the dog handlers from the dog shelters had poor knowledge (90%, n=34/38) and attitude (100%, n=38/38) and moderate practice (53%, n=20/38) towards leptospirosis, whereas the working dog handlers had poor knowledge (58%, n=58/100) and attitude (75%, n=75/100) but a good level of practice (77%, n=77/100). The poor attitude towards leptospirosis in both groups is alarming as it may hinder implementation plans of control and preventive measures. Although there was good hand-washing practices among the dog handlers, there was a lack of personal protective equipment (PPE) practice that may therefore expose both the dogs and dog handlers to leptospirosis. Information gained from this study could be used to implement intervention programmes and to suggest prevention measures to limit the risk of leptospirosis among the dog handlers.
  17. Lee SV, Tai ES, Mutalib AR, Khairani-Bejo S, Bahaman AR
    Trop Biomed, 2011 Dec;28(3):497-505.
    PMID: 22433877 MyJurnal
    A simple and reliable tool for the early diagnosis of leptospirosis is urgently needed. We report the development of a lyophilized reagent-based polymerase chain reaction (PCR) assay targeting lipL32 gene, which is present only in pathogenic leptospires. To determine the effectiveness of the newly developed assay in the early diagnosis of leptospirosis, the sensitivity and specificity was evaluated. In simulated clinical samples, the assay was able to detect 10² and 10³ leptospires/ml in spiked urine and blood samples, respectively. In experimentally infected animals, leptospiral DNA could be detected in blood and lung samples as early as Day 1 post infection. This assay was also shown to be stable and remained sensitive for up to five months at ambient temperature. Hence, this lyophilized reagent-based PCR assay with high specificity, sensitivity and stability would provide a simple, rapid and reliable method in diagnosing acute leptospirosis, especially in the field of veterinary medicine.
  18. Mazlan M, Khairani-Bejo S, Hamzah H, Nasruddin NS, Salleh A, Zamri-Saad M
    Vet Q, 2021 Dec;41(1):36-49.
    PMID: 33349157 DOI: 10.1080/01652176.2020.1867328
    BACKGROUND: Brucellosis 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.

    OBJECTIVE: To describe the pathological changes, identification and distribution of B. melitensis in foetuses of experimentally infected does.

    METHODS: Twelve 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.

    RESULTS: Bilateral 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.

    CONCLUSION: Vertical 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.

  19. Mazlina M, Khairani-Bejo S, Hazilawati H, Shaqinah NN, Zamri-Saad M
    Transbound Emerg Dis, 2021 Jul;68(4):2028-2038.
    PMID: 32979887 DOI: 10.1111/tbed.13850
    This study describes the pathological changes, antibody response, isolation and distribution patterns following exposure of non-pregnant goats to live Brucella melitensis. Eighteen healthy adult female goats were divided into two equal groups. Group 1 was infected via conjunctival sac with 109  cfu/ml of B. melitensis, while Group 2 was similarly exposed to sterile PBS. Serum and swabs from the eyes and vagina were collected at 5-day intervals. On days 15, 30 and 75 post-infection, 3 goats from each group were killed before the conjunctiva, ovary, oviduct, uterine horn, uterine body and vagina, the submandibular, prescapular and supramammary lymph nodes, the mammary gland, liver, spleen, urinary bladder and synovial membranes were collected for bacterial isolation and pathological study. Exposure of non-pregnant goats to B. melitensis did not produce clinical signs and gross lesions but produced mild necrosis and inflammation in the lymph nodes, the organs of reproductive tract, the mammary gland and urinary bladder. In general, microscopic lesions were most severe in the D75 goats, followed by D30 and D15 goats. Brucella melitensis was most frequent and significantly (p 
  20. Mazlina M, Khairani-Bejo S, Hazilawati H, Tiagarahan T, Shaqinah NN, Zamri-Saad M
    BMC Vet Res, 2018 Jun 25;14(1):203.
    PMID: 29940976 DOI: 10.1186/s12917-018-1533-x
    BACKGROUND: This study was conducted to investigate the pathological changes and distribution of B. melitensis in the urinary tract of pregnant goats following acute experimental infection. Six Jamnapari crossbred does in their third trimester of pregnancy were randomly assigned into two groups; Group 1 was uninfected control and Group 2 was inoculated conjunctival with 0.1 mL of the inoculums containing 109 cfu/mL of live B. melitensis. All does were sacrificed 30 days post-inoculation before the kidney, ureter, urinary bladder, urethra and vaginal swab were collected for isolation of B. melitensis. The same tissue samples were fixed in 10% neutral buffered formalin for hematoxylin and eosin, and immunoperoxidase staining.

    RESULTS: None of the goats showed clinical signs or gross lesions. The most consistent histopathology finding was the infiltration of mononuclear cells, chiefly the macrophages with few lymphocytes and occasionally neutrophils in all organs along the urinary tract of the infected goats of Group 2. Other histopathology findings included mild necrosis of the epithelial cells of the renal tubules, congestion and occasional haemorrhages in the various tissues. Kidneys showed the most severe lesions. Immunoperoxidase staining revealed the presence of B. melitensis within the infiltrating macrophages and the epithelium of renal tubules, ureter, urethra and urinary bladder. Most extensive distribution was observed in the urinary bladder. Brucella melitensis was successfully isolated at low concentration (3.4 × 103 cfu/g) in the various organs of the urinary tract and at high concentration (2.4 × 108 cfu/mL) in the vaginal swabs of all infected goats. Although B. melitensis was successfully isolated from the various organs of the urinary tract, it was not isolated from the urine samples that were collected from the urinary bladder at necropsy.

    CONCLUSION: This study demonstrates the presence of low concentrations of B. melitensis in the organs of urinary tract of pregnant does, resulting in mild histopathology lesions. However, B. melitensis was not isolated from the urine that was collected from the urinary bladder.

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