The perspective of antibiotic therapeutic challenges of brucellosis in the Middle East and North African countries: Current situation and therapeutic management.

Brucellosis is among the most prevalent zoonotic infections in Middle Eastern and North African (MENA) countries, critically impacting human and animal health. A comprehensive review of studies on antibiotic susceptibility and therapeutic regimes for brucellosis in ruminants and humans in the MENA region was conducted to evaluate the current therapeutic management in this region. Different scientific databases were searched for peer-reviewed original English articles published from January 1989 to February 2021. Reports from research organizations and health authorities have been taken into consideration. Brucella melitensis and Brucella abortus have been reported from the majority of MENA countries, suggesting a massive prevalence particularly of B. melitensis across these countries. Several sporadic cases of brucellosis relapse, therapeutic failure, and antibiotic resistance of animal and human isolates have been reported from the MENA region. However, several studies proved that brucellae are still in-vitro susceptible to the majority of antibiotic compounds and combinations in current recommended World Health Organization (WHO) treatment regimens, for example, levofloxacin, tetracyclines, doxycycline, streptomycin, ciprofloxacin, chloramphenicol, gentamicin, tigecycline, and trimethoprim/sulfamethoxazole. The current review presents an overview on resistance development of brucellae and highlights the current knowledge on effective antibiotics regimens for treating human brucellosis.

Spatial variation in direct and indirect contact rates at the wildlife-livestock interface for informing disease management.

Little is known about disease transmission relevant contact rates at the wildlife-livestock interface and the factors shaping them. Indirect contact via shared resources is thought to be important but remains unquantified in most systems, making it challenging to evaluate the impact of livestock management practices on contact networks. Free-ranging wild pigs (Sus scrofa) in North America are an invasive, socially-structured species with an expanding distribution that pose a threat to livestock health given their potential to transmit numerous livestock diseases, such as pseudorabies, brucellosis, trichinellosis, and echinococcosis, among many others. Our objective in this study was to quantify the spatial variations in direct and indirect contact rates among wild pigs and cattle on a commercial cow-calf operation in Florida, USA. Using GPS data from 20 wild pigs and 11 cattle and a continuous-time movement model, we extracted three types of spatial contacts between wild pigs and cattle, including direct contact, indirect contact in the pastoral environment (unknown naturally occurring resources), and indirect contact via anthropogenic cattle resources (feed supplements and water supply troughs). We examined the effects of sex, spatial proximity, and cattle supplement availability on contact rates at the species level and characterized wild pig usage of cattle supplements. Our results suggested daily pig-cattle direct contacts occurred only occasionally, while a significant number of pig-cattle indirect contacts occurred via natural resources distributed heterogeneously across the landscape. At cattle supplements, more indirect contacts occurred at liquid molasses than water troughs or molasses-mineral block tubs due to higher visitation rates by wild pigs. Our results can be directly used for parameterizing epidemiological models to inform risk assessment and optimal control strategies for controlling transmission of shared diseases.

Socio-economic impacts of brucellosis on livestock production and reproduction performance in Koibatek and Marigat regions, Baringo County, Kenya.

BACKGROUND: Brucellosis in Africa is caused by Brucella species transmitted through contaminated or contacts with infected animals or their carcasses. The disease reduces livestock production and reproduction performance evident by frequent episodes of abortion, still births, swollen testes, weak calves/lambs and swollen joints. However, the socio-economic impacts of these brucellosis-associated symptoms on milk, fat, meat and blood production, infertility, sale value, dowry and costs of treatment has not been evaluated extensively in developing countries. In Baringo County, Kenya, there is a continuous movement of cattle as a result of trade and grazing, which predisposes many herds to brucellosis infection. The objective of this study was to investigate the socio-economic impacts of Brucella infection on production systems for sheep, goats, cattle and camels and explore the impact of brucellosis on livestock production and reproduction performance among livestock keeping communities in Baringo County, Kenya. The study adopted a cross-sectional survey using quantitative data collection methods. RESULTS: Results demonstrated an impact on milk production in suspected brucellosis cases resulting from abortions (OR = 0.151, P < 0.0001) and swollen joints (OR = 2.881, P < 0.0001). In terms of infertility, abortion as a symptom of brucellosis (OR = 0.440, P = 0.002), still birth (OR = 0.628, P = 0.042), and weak calf or lamb (OR = 0.525, P = 0.005) had an impact on infertility. In terms of sale value, abortion (OR = 0.385, P = 0.008), weak calf/lamb (OR = 2.963, P = 0.013) had an impact on sale value. Other analyses demonstrated that for dowry, swollen testes (OR = 5.351, P = 0.032), weak calf and lambs (OR = 0.364, P = 0.019) had a likelihood of reduction of dowry value. Finally, in terms of cost of treatment, abortion (OR = 0.449, P = 0.001), still births (OR = 0.208, P = 0.015), swollen testes (OR = 0.78, P = 0.014), weak calf/lambs (OR = 0.178, P = 0.007) and swollen joints (OR = 0.217, P = 0.003) significantly increased the costs of treatments. There was no impact on fat and meat and blood production. CONCLUSION: Even though there was a huge socio-economic impact on milk production, infertility, sale value, and dowry, it was the costs of treatment that was significantly impacted on all symptoms associated with brucellosis on this community. A 'One Health' approach in tackling the brucellosis menace as a holistic approach is recommended for both humans and their livestock.

Antimicrobial Resistance in Leptospira, Brucella, and Other Rarely Investigated Veterinary and Zoonotic Pathogens.

Leptospira, Brucella, and Borrelia are major agents of zoonotic disease, causing high morbidity and, in some cases, significant mortality in humans. For all three genera, prompt diagnosis and appropriate antimicrobial therapy are required to prevent the development of chronic, debilitating illness. Leptospira spp. are intrinsically resistant to several antimicrobial classes; however, there is little evidence in the literature for development of acquired resistance to antimicrobial agents used for clinical treatment of acute leptospirosis. For Brucella infections, there are numerous reports of relapses following therapy, but it is unclear whether this is due to sequestration within infected sites (e.g., bone) or the development of acquired resistance. Brucella have maintained their susceptibility to doxycycline and rifampicin, which in combination remain the most common treatments of brucellosis in humans. In vitro induced point mutations are described as imparting resistance to rifampicin (rpoB) and fluoroquinolones (gyrA). The clinical significance of these mutations is unclear. For Borrelia burgdorferi, although acquired resistance to some antimicrobial agents has been described, resistance due to bacterial persister cells surviving in the presence of antimicrobial, with no apparent increase in the MIC of the organism, have been recently described. Of the remaining veterinary fastidious pathogens, Lawsonia intracellularis is the most interesting from an antimicrobial resistance perspective because it can only be grown in cell culture, making in vitro susceptibility testing challenging. MIC testing has been undertaken on a small number of isolates, and some differences in susceptibility to macrolides have been demonstrated between isolates obtained from different regions.

Inhibitory effect of red ginseng acidic polysaccharide from Korean red ginseng on phagocytic activity and intracellular replication of Brucella abortus in RAW 264.7 cells.

Korean red ginseng (KRG) has long been used in traditional Korean and Oriental medicine. However, the anti-bacterial mechanism and therapeutic efficiency of KGR for intracellular Brucella infection are still unclear. In this study, the bactericidal activity of Korean red ginseng acidic polysaccharide (RGAP) on Brucella (B.) abortus and its cytotoxic effects on RAW 264.7 cells were evaluated. In addition, B. abortus internalization and intracellular replication in macrophages were investigated after RGAP treatment. RGAP-incubated cells displayed a marked reduction in the adherence, internalization and intracellular growth of B. abortus in macrophages. Furthermore, decreased F-actin fluorescence was observed relative to untreated B. abortus-infected cells. Western blot analysis of intracellular signaling proteins revealed reduced ERK, JNK and p38α phosphorylation levels in B. abortus-infected RGAP-treated cells compared to the control. Moreover, elevated co-localization of B. abortus-containing phagosomes with lysosome-associated membrane protein 1 (LAMP-1) were observed in RGAP-treated cells compared with the control. Overall, the results of this study suggest that RGAP can disrupt phagocytic activity of B. abortus via suppression of mitogen-activated protein kinases (MAPKs) signaling proteins ERK, JNK and p38 levels and inhibit intracellular replication of B. abortus by enhancing phagolysosome fusion, which may provide an alternative control of brucellosis.

Studies on a suitable antibiotic therapy for treating swine brucellosis.

The aim of this work was developing effective treatments against Brucella suis biovar 2, responsible for swine brucellosis in Europe. MICs for antibiotics used classically in brucellosis and two new macrolides (tulathromycin and tildipirosin) were determined for 33 B. suis biovar 2 field and B. suis reference strains. MIC90 values ranged from 0.01 to 0.25 µg/mL. The best candidates, given alone or combined, were then evaluated in mice. Ten groups (n = 7) of BALB/c mice were inoculated (1 × 10(5) CFU/mouse) with a virulent B. suis biovar 2 field strain. All groups, excepting untreated control, were treated for 14 days with, respectively, doxycycline, dihydrostreptomycin, tulathromycin (one or two doses), or tildipirosin (one or two doses) given alone, and doxycycline combined with dihydrostreptomycin, tulathromycin, or tildipirosin. Combined tildipirosin treatment was the most effective, then selected for pig studies. Sixteen B. suis biovar 2 naturally infected sows were treated with oxytetracycline (20 mg/kg BW/daily) for 21 days. The half of these received also tildipirosin (4 mg/kg BW) in two doses with a 10-day interval. An extensive bacteriological study conducted ten days after ceasing treatments proved the efficacy of this combined oxytetracycline/tildipirosin treatment.

Brucellosis in domestic water buffalo (Bubalus bubalis) of Trinidad and Tobago with comparative epidemiology to cattle.

The water buffalo is an important domestic animal worldwide, and the local Buffalypso variety was developed in Trinidad to have improved beef qualities. Brucellosis was diagnosed in Trinidad and Tobago during 1998 in both cattle and domestic water buffalo (Bubalus bubalis) populations. Brucellosis in the latter species is caused by infection with Brucella abortus, similar to bovine brucellosis. Control of brucellosis is of paramount importance to preservation of the genetic diversity of these animals in Trinidad, and this has been complicated by differences in the epidemiology of water buffalo and bovine brucellosis. Some diagnostic tests do not have comparable accuracy between the two species, and the RB51 vaccine does not adequately protect against infection in water buffalo. The water buffalo in Trinidad may also be more resistant to infection than cattle. Development of effective vaccination protocols is key to brucellosis control in Buffalypso in Trinidad, and prohibitions on import of virulent B. abortus strains for vaccine efficacy studies has impeded progress in this area. These Trinidadian strains are of variable virulence; some might be effective for challenge in vaccine efficacy studies, while other, of lower virulence, may be vaccine candidates for use in water buffalo.

Use of enrofloxacin in the treatment of canine brucellosis in a dog kennel (clinical trial).

To date, no totally effective antibiotic for the eradication of canine brucellosis has been found. The purpose of this study was to evaluate the efficacy of enrofloxacin in a kennel infected with Brucella canis. Twelve dogs, 2 males and 10 females (including 1 in estrus, 3 pregnant, and 6 in anestrus) infected with B. canis were given 5 mg/kg of enrofloxacin orally every 12 h for 30 days. Females received additional courses of enrofloxacin during the estral and luteal phases of the subsequent cycles (0-2 cycles). They were repeatedly mated by infected males. A serological follow-up was carried out for 38 months. The clinical, serological and bacteriological findings were recorded. In a trial carried out 14 months after the beginning of this study, all dogs were negative on the Rapid Slide Agglutination Test (RSAT). No abortions were observed. All mated female dogs conceived and gave birth to healthy puppies. Cultures of postpartum vaginal discharges (lochia) were negative for B. canis. Similar to other treatments, although enrofloxacin was not completely efficacious in treating canine brucellosis, it maintained fertility and avoided the recurrence of abortions, transmission of the disease to the puppies and dissemination of microorganisms during parturition. We inferred that enrofloxacin could be used as an alternative drug for the treatment of canine brucellosis.

Serological responses of rams to a Brucella ovis-vitamin E adjuvant vaccine.

Rams which were vaccinated at 6-8 months of age with a water-in-oil Brucella ovis-vitamin E adjuvant vaccine had significantly higher serum antibody levels than rams vaccinated with a commercial B. ovis bacterin or B. melitensis Rev 1. The adjuvant vaccine did not cause abscesses at the site of injection as some water-in-oil emulsions do. Two years after vaccination, the vitamin E adjuvant-vaccinated rams had higher antibody level than the other groups. This was most likely due to a secondary response to naturally occurring infection with B. ovis.

Experimental treatment of Brucella melitensis infection in sheep with oxytetracycline alone or combined with streptomycin.

The efficacy of oxytetracycline (OTC) alone or combined with streptomycin in the treatment of 118 Najdi ewes believed to have been naturally infected with Brucella melitensis, was evaluated by culture of selected tissues and organs at slaughter. Groups of sheep were given 250, 500 or 1,000 mg of OTC intraperitoneally (i/p) daily for six weeks and in the respective groups at necropsy 52, 69 and 100% of sheep were found to be Brucella-free. Treatment with 250 mg OTC (daily for six weeks i/p) combined with 1,000 mg streptomycin (daily for three weeks intramuscularly) increased the percentage of Brucella-free sheep to 82%. When a group of sheep were each inoculated i/p with 1,000 mg of long-acting OTC every three days over a period of six weeks, 75% of them were Brucella-free at necropsy. B. melitensis was isolated from all (24) non-treated (control) sheep. The results showed that long-term treatment with a high dose of OTC alone had succeeded in eliminating B. melitensis from a group of 16 naturally infected sheep.

Protection of rams against epididymitis by a Brucella ovis-vitamin E adjuvant vaccine.

Rams vaccinated at 7 and 8 months of age with a B. ovis-vitamin E adjuvant vaccine had increased antibody titers compared with Freund's incomplete adjuvant or commercial bacterin vaccinated rams. The percent overall infectivity in an experimental infection of B. ovis-vitamin E adjuvant vaccinated rams was 22% compared to 44% for B. ovis-Freund's incomplete adjuvant or bacterin vaccinated rams and 67% for control.

Brucella abortus vaccines: comparison of protection provided by immunopotentiated 45/20 bacterins and live strain 19 vaccine in guinea pigs.

Guinea pigs were subcutaneously inoculated with 300 microgram of Brucella abortus strain 45/20 killed cells combined in 1% oil emulsion with trehalose dimycolate (TDM), muramyl dipeptide (MDP), or a combination of the 2 immunopotentiators. Protection, as determined by splenic infections in the guinea pigs after challenge exposure, was compared with that induced by strain 19 vaccine. With few exceptions, protection induced by bacterins containing 50 to 1,000 microgram of TDM or TDM-MDP/dose was comparable with that of strain 19 vaccine (P greater than 0.05). Bacterins that contained MDP as an adjuvant were inferior to those with TDM regardless of the excipient or method of preparation. There was no further enhancement of immunogenicity by the addition of MDP to bacterins that already contained TDM. Mineral oil could not be replaced by a metabolizable excipient in bacterins potentiated with TDM.

Symposium on back problems in the horse. (2) The diagnosis of diseases of the horse's back.

A description of some of the clinical features of low back pain in the horse has been given and a number of methods for assisting diagnosis considered. As well as a complete clinical examination both at rest and during exercise, a useful diagnostic aid in some chronic cases was the injection of local anaesthetic into the interspinous spaces. A laboratory examination, including haematological and biochemical profiles, was undertaken in all cases. The serum enzymes GOT and CPK were particularly valuable as an aid to diagnosis in atypical tying-up. A technique for radiography of the vertebral column of the mid back in the standing position and of the pelvic and sacroiliac regions of the anaesthetised horse was described using a Siemens Triplex Optimatic 1023 machine. Some of the radiological features of the vertebral column were considered and a breakdown of the diagnosis of 110 referred back cases. The most important conditions included muscle strain, crowding and overriding of the dorsal spinous processes in the mid back, spondylosis, undue curvature of the spine and vertebral fractures.