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Background: Several diagnostic environments in Uganda lack real-time, robust and high-throughput technologies for comprehensive typing of microbes, which is a setback to infectious disease surveillance. This study combined various wet laboratory diagnostics to understand the epidemiology of pathogenic staphylococci isolated from animals in Uganda and the implications for global health security priorities. Methods: A retrospective study was conducted employing records and pathogenic staphylococci (from animals) archived at the Central Diagnostic Laboratory (CDL), Makerere University, Uganda, between January 2012 and December 2019. The bacteria were speciated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and tested for virulence factors [beta lactamases, lecithinase, deoxyribonuclease (DNase), haemolysins] and resistance to ten antimicrobials of clinical and veterinary relevance. Tetracycline and methicillin resistance genes were also tested. Results: The prevalent diseases were mastitis in cattle and skin infections in dogs. Of the 111 staphylococci tested by MALDI-TOF MS, 79 (71.2%) were Staphylococcus aureus, 27 (24.3%) were Staphylococcus pseudintermedius and 5 (4.5%) were Staphylococcus schleiferi. All these strains expressed haemolysins. The prevalence of strains with lecithinase, penicillinase, cephalosporinase and DNase was 35.9% (14/39), 89.7% (35/39), 0.0% (0/39) and 87.2% (34/39), respectively. Staphylococci were primarily resistant to early penicillins (over 80%), tetracycline (57.7%), and chloramphenicol (46.2%). Minimal resistance was noted with cloxacillin (0.0%), ciprofloxacin (9.6%), and cefoxitin (3.8%). The prevalence of multidrug resistance (MDR) was 78.8% for general staphylococci, 82.2% for S. aureus, 73.1% for S. pseudintermedius, and 60.0% for S. schleiferi. Multidrug resistant staphylococci were significantly more prevalent in the cattle isolates than in the dog isolates (P < 0.05). The prevalence of methicillin-resistant staphylococci (MRS) tested by resistance to cefoxitin and mecA carriage was 3.8%. These four strains were all isolated from dog skin infections. The tetK gene was the most predominant (35.4%), followed by tetM (25.0%). Conclusion: In resource-constrained settings, the approach of integrated diagnostics promises sustainable disease surveillance and the addressing of current capacity gaps. The emergence of MRS (zoonotic bacteria) in companion animals creates a likelihood of reduced treatment options for related human infections, a threat to global health. [ABSTRACT FROM AUTHOR]

Background: Mastitis and associated antimicrobial resistance (AMR) are major challenges to the dairy industry worldwide. Objective: This study aimed to expose the mastitis burden, causative bacteria and drivers for mastitis‐causing multi‐drug‐resistant (MDR) Staphylococci infectivity in cows on dairy farms in Wakiso district, Uganda. Methods: On 22 farms, practices were documented using questionnaires, and 175 cows were screened by the California mastitis test. Composite milk samples from the positive reactors were submitted to the laboratory for bacterial culture testing. Antimicrobial sensitivity testing by the Kirby Bauer disc diffusion method was done only on Staphylococci with a panel of 10 antimicrobials of clinical relevance. Results: Mastitis was detected in 80.6% (n = 141) of the 175 sampled cows, of which sub‐clinical mastitis (76.0%: n = 133) was predominant. The Chi‐squared analysis hypothesized that cow age (p = 0.017), sub‐county (p = 0.013), parity (p < 0.0001), sex of farm owner (p = 0.003), farm duration in dairy production (p = 0.048) and the use of milking salve (p = 0.006) were associated with mastitis. Coagulase‐negative Staphylococci were the most prevalent (71.4%; n = 95), followed by Staphylococcus aureus (30.1%, n = 40). Staphylococci (76.3%; n = 135) were majorly resistant to penicillin and tetracycline. Only one isolate was phenotyped as a methicillin‐resistant Staphylococcus specie (MRSS). The prevalences of MDR strains at cow and isolate level were 6.3% and 8.3%. The major MDR phenotype identified was penicillin–tetracycline–trimethoprim‐sulphamethoxazole. The isolate detected as an MRSS exhibited the broadest MDR pattern. Cow parity was identified as a predictor of infectivity of mastitis‐causing MDR Staphylococci in dairy herds. Conclusion: The high prevalence of mastitis and associated pathogen AMR found exposes possibilities of economic losses for the dairy sector warranting the need for farmer sensitization on the institution of proper mastitis prevention and control programs, with emphasis on milking hygiene practices and routine disease monitoring. [ABSTRACT FROM AUTHOR]

We conducted a cross-sectional serologic study at Kampala City abattoir in Uganda on 287 small ruminants (221 goats and 66 sheep) to determine the seroprevalence of brucellosis. The samples were tested using a modified rose bengal test (mRBT) and an indirect ELISA (iELISA). Small ruminant Brucella spp. seropositivity was 18 of 287 (6.3%) by mRBT and 19 of 287 (6.6%) by iELISA. The prevalence of brucellosis by mRBT was non-significantly higher in goats (17 of 221; 7.7%) than in sheep (1 of 66, 1.5%; p = 0.069), and also non-significantly higher by the iELISA in goats (18 of 221; 8.1%) than in sheep (1 of 66, 1.5%; p = 0.057). Brucellosis in slaughtered goats and sheep is a public health hazard to abattoir workers and consumers that calls for control and eradication measures at the farm level, given that testing is not carried out routinely at slaughter points. [ABSTRACT FROM AUTHOR]

The dynamics of extended-spectrum β-lactamase- (ESBL-) and AmpC β-lactamase-producing bacteria (which are deadly groups of antimicrobial-resistant bacteria) have not been well understood in developing countries. This raises major concerns to antimicrobial resistance (AMR) control. We investigated the prevalence and factors linked to the fecal carriage of ESBL- or AmpC-producing Escherichia coli (ESBL-/AmpC-EC) in commercial chickens. Cloacal swabs from 400 birds were sampled and submitted to the Central Diagnostic Laboratory for ESBL-/AmpC-EC screening by culture methods using MacConkey agar supplemented with cefotaxime. Epidemiological data were collected using a structured questionnaire and plausible risk factor analyses prepared by R software using X2 test and logistic regression modeling. Results showed that the prevalence of ESBL-/AmpC-EC was 17.5%. Univariable screening hypothesized that carriage was probably influenced by a type of commercial chicken, geographical location, age group, flock size, and housing system (p < 0.05). Modeling exposed that broiler birds were at a higher risk of being ESBL-/AmpC-EC carriers (COR = 9.82, CI = 3.85–25.07). Birds from Wakiso Town Council (COR = 4.89, CI = 2.04–11.72) and flocks of 700–1200 birds were also at a higher risk of harboring ESBL-/AmpC-EC (COR = 2.41, CI = 1.11–5.23). Birds aged 4 months and below were more susceptible to ESBL-/AmpC-EC carriage compared with those aged 1 month and below being 6.33 times (CI = 1.65–24.35) likely to be carriers. The occurrence of ESBL-/AmpC-EC in flocks suggests possible treatment failures while managing colibacillosis. Consequently, injudicious antimicrobial use should be replaced with an accurate diagnosis by bacterial culture and sensitivity testing so as to circumvent AMR emergence, spread, and associated losses. [ABSTRACT FROM AUTHOR]

Mycoplasmas are the smallest free-living organisms and cause a number of economically important diseases affecting humans, animals, insects, and plants. Here, we demonstrate that highly virulent Mycoplasma mycoides subspecies capri (Mmc) can be fully attenuated via targeted deletion of non-essential genes encoding, among others, potential virulence traits. Five genomic regions, representing approximately 10% of the original Mmc genome, were successively deleted using Saccharomyces cerevisiae as an engineering platform. Specifically, a total of 68 genes out of the 432 genes verified to be individually non-essential in the JCVI-Syn3.0 minimal cell, were excised from the genome. In vitro characterization showed that this mutant was similar to its parental strain in terms of its doubling time, even though 10% of the genome content were removed. A novel in vivo challenge model in goats revealed that the wild-type parental strain caused marked necrotizing inflammation at the site of inoculation, septicemia and all animals reached endpoint criteria within 6 days after experimental infection. This is in contrast to the mutant strain, which caused no clinical signs nor pathomorphological lesions. These results highlight, for the first time, the rational design, construction and complete attenuation of a Mycoplasma strain via synthetic genomics tools. Trait addition using the yeast-based genome engineering platform and subsequent in vitro or in vivo trials employing the Mycoplasma chassis will allow us to dissect the role of individual candidate Mycoplasma virulence factors and lead the way for the development of an attenuated designer vaccine. [ABSTRACT FROM AUTHOR]

Mastitis and antimicrobial resistance are a big challenge to the dairy industry in sub-Saharan Africa. A study was conducted in Kashongi and Keshunga subcounties of Kiruhura District (in Uganda) where the government and private sector have deliberate programs to improve production efficiency, quality, and safety of milk and its products. The study aimed to determine the prevalence of mastitis, its common causative agents, antimicrobial sensitivity of mastitis causing organisms, and contaminants of processed milk products: yoghurt and ghee. Seventy-one milk, fourteen yoghurt, and three ghee samples were collected from nine farms. Of the 71 cows tested, 54 (76.1%) had mastitis. The mastitis cases from Keshunga were 32 (59.3%) and Kashongi contributed 22 (40.7%) of the cases. The common mastitis causative agents were Staphylococcus spp. (30.8%), Streptococcus spp. (12.3%), Corynebacterium spp.(15.4%), and E. coli (7.7%). Some of the isolates were resistant to tetracycline and penicillin. Prevalent contaminants of yoghurt were Staphylococcus spp. (8.3%), Streptococcus spp. (8.3%), Corynebacterium spp. (8.3%), and E. coli (8.3%), whereas all ghee contained Streptococcus spp. (100%). Prevalence of mastitis, antimicrobial resistance, and contamination of milk products are high in the study area. Targeted programs to prevent and control mastitis as well as antibiotic resistance are recommended. [ABSTRACT FROM AUTHOR]