Non-typhoidal Salmonella contributes to gastrointestinal infections in Morogoro: Evidence from patients attending Morogoro regional referral hospital in Tanzania.

INTRODUCTION: Salmonella is one of the most common causes of food-borne outbreaks and infection worldwide. Non-typhoidal Salmonella (NTS) infections are common and remain a significant public health problem among important bacterial foodborne diseases. The current study aimed to establish the Non typhoidal Salmonella infection and antimicrobial resistance status among selected patients at Morogoro Regional Referral Hospital (MRRH), Morogoro Region, Tanzania, to inform clinical care management and public health interventions. METHODOLOGY: A cross-sectional study was conducted using medical records and samples were collected from hospitalised and outpatients between October and December 2021. A total of 153 participants were enrolled in the study and 132 consented to being sampled. The collected samples were analysed using standard microbiological techniques. The isolates were subjected to molecular genotyping, where Polymerase Chain Reaction (PCR) was performed targeting the 16S rDNA gene. PCR products were then submitted for sequencing to establish phylogenetic relatedness. Antimicrobial susceptibility testing and resistance genes screening were also conducted. RESULTS: The phylogenetic analysis identified two Salmonella serovars; Salmonella Enteritidis and Salmonella Typhimurium. The isolates were from four adults and seven children patients. The isolates were tested against six antimicrobial agents: tetracycline, trimethoprim, gentamycin, ciprofloxacin, ampicillin and cefotaxime. Further antimicrobial assays were performed by screening 10 antimicrobial resistance genes using PCR. Overall, the highest resistance was observed in ampicillin (100%), whereas the lowest resistance was recorded for ciprofloxacin and gentamicin (9.1%). In addition, four (36.4%) of the isolates were resistant to cefotaxime and three (27.3%) to tetracycline and trimethoprim. The isolates also exhibit the presence of resistance genes for sulfamethoxazole 1&2, tetracycline (tet) A&B, Beta-lactamase CTXM, Beta-lactamase TEM, Beta-lactamase SHV, Gentamycine, Acra and acc3-1 in different occurrences. The overall prevalence of Salmonella species in Morogoro region was 8.3% (11/132) with Salmonella Enteritidis and Salmonella Typhimurium being the only serovars detected from adults and children stool samples. CONCLUSION: Our investigation showed that both children and adults had been exposed to Salmonella spp. However, the occurrence of NTS was higher in children (5.3% (7/132) compared to adults (3.0% (4/132). To stop zoonotic infections and the development of antimicrobial resistance in the community, this calls for Infection Prevention and Control (IPC) and stewardship programmes on rational use of antimicrobials in both health facilities and at the community level.

Mitigating antimicrobial resistance (AMR) using implementation research: a development funder's approach.

Despite the escalating burden of antimicrobial resistance (AMR), the global response has not sufficiently matched the scale and scope of the issue, especially in low- and middle-income countries (LMICs). While many countries have adopted national action plans to combat AMR, their implementation has lagged due to resource constraints, dysfunctional multisectoral coordination mechanisms and, importantly, an under-recognized lack of technical capacity to adapt evidence-based AMR mitigation interventions to local contexts. AMR interventions should be tailored, context-specific, cost-effective and sustainable. The implementation and subsequent scale-up of these interventions require multidisciplinary intervention-implementation research (IIR). IIR involves both quantitative and qualitative approaches, occurs across a three-phase continuum (proof of concept, proof of implementation and informing scale-up), and across four context domains (inner setting, outer setting, stakeholders and the implementation process). We describe the theoretical underpinnings of implementation research (IR), its various components, and how to construct different IR strategies to facilitate sustainable uptake of AMR interventions. Additionally, we provide real-world examples of AMR strategies and interventions to demonstrate these principles in practice. IR provides a practical framework to implement evidence-based and sustainable AMR mitigation interventions.

Existence of a novel heavy metal-tolerant Pseudomonas aeruginosa strain Zambia SZK-17 Kabwe 1: the potential bioremediation agent in the heavy metal-contaminated area.

Bacterial biomass may serve as an important environmental cleaning agent to toxic heavy metal ions at the expense of chemical processes which are not environmentally friendly. This study aimed at characterizing bacterial agents which could serve as a potential in situ bioremediation agent at the site of isolation. The characterization was performed using both phenotypic and molecular approaches. A novel Pseudomonas aeruginosa strain Pseudomonas aeruginosa Zambia SZK17 Kabwe1 was successfully isolated, identified, and characterized. The strain showed a promising tolerance to heavy metals such as copper (2 mM), zinc, nickel (2 mM), cobalt (1 mM), and cadmium (0.5 mM) at the laboratory level. The bacterium has shown the bioaccumulation of at least 60% of copper (II) sulfate (0.3655 mg/l) with R = 69.75%, cadmium (II) chloride (0.0241 mg/l) with R = 69.98%, zinc (II) chloride (0.1389 mg/l) with R = 69.91%, nickel (II) chloride (0.1155 mg/l) with R = 69.92%, and cobalt (II) chloride (0.593 mg/l) with R = 69.92%. The highest bioaccumulation has been observed in heavy metals cadmium, zinc, nickel, and cobalt. Characterization of the bacterium on pH has revealed that at a very high pH (≥ 9) and lower (≤ 5.5) pH, the bacterium tended to have reduced growth with optimum growth at pH 8. The high temperature at around 40 °C had a negative effect on the growth performance of the bacterium while optimum growth was observed at 28 °C. This novel P. aeruginosa strain has shown the phenotypic attributes to become a potential bioremediation agent; however, further investigation needs to be done to understand the genes and or molecular mechanisms that drive their tolerance to multiple heavy metals.

Phenotypic and molecular detection of Aeromonads infection in farmed Nile tilapia in Southern highland and Northern Tanzania.

Aeromonads disease outbreaks are now becoming a common phenomenon in freshwater farmed fish worldwide. In Tanzania, the aquaculture field is increasingly growing save to sustain food protein demand and strengthen household income. To avoid losses that tilapia fish farmers might account, information on magnitude of infection and characteristics of the aetiological agent is vital. This study aimed to establish the prevalence of aeromonads infection in farmed tilapia and assess pond and fish health management practices. A cross sectional study was carried out between February 2017 and October 2018 and a total of 816 whole fish samples were aseptically collected from 32 ponds in Ruvuma, Mbeya, Iringa and Kilimanjaro regions. During sampling, water quality parameters were taken and questionnaires to assess the knowledge of farmers were also provided. Isolation and identification of bacteria was conducted using conventional biotyping and molecular techniques. A total of 201 (80.4%) of 250 isolates that were conventionally identified were confirmed to be aeromonads by amplification of 820 bp rpoD gene, making the overall prevalence of 24.6% (201, n = 816). Sequencing of rpoD gene and phylogenetic analysis revealed two aeromonads species, Aeromonas hydrophila and Aeromonas veronii. To the best of our knowledge this is the first report to establish the prevalence of aeromonads in apparently healthy farmed tilapia in Southern highlands and Northern zone of Tanzania. In addition it was observed that farmers were lacking proper knowledge and awareness on pond management practices and fish health management. In conclusion, the infection rate of aeromonads in apparently health tilapia coupled with lack of proper knowledge and awareness on pond and fish health management by fish farmers in the study area poses risk of diseases outbreaks in their farms in future. Therefore, it is recommended that the farmers should be trained on basic pond and fish health management and control strategies.

Current State of Modern Biotechnological-Based Aeromonas hydrophila Vaccines for Aquaculture: A Systematic Review.

This systematic review describes what "the cutting edge vaccines for Aeromonas hydrophila are". The focus is on types of high tech biotechnological based vaccines, target gene or antigen in developing these vaccines, and challenge model fish species used in vaccines efficacy testing. Vaccines delivery methods, immune response, and their efficacy, adjuvant or carrier systems used, and the overall experimental setup or design of the vaccines under investigation are also described. The search for the original papers published between 2009 and 2018 was conducted in June of 2018, using the PubMed and Google scholar electronic database. Twenty-three (23/4386) studies were included in the final assembly using PRISMA guidelines (Protocol not registered). Recombinant protein vaccines were the highly experimented type of the modern biotechnological based vaccines identified in the selected studies (16/23; 70%). Outer membrane proteins (OMPs) of different ß-barrels were shown to be a potential antigenic entity for A. hydrophila vaccines (57%). Intraperitoneal route with conventional carries or adjuvants was the highly applied delivery system while very few studies used herbal based vaccine adjuvants and nanomaterial as a vaccine carrier. Variation was observed in terms of protection levels in the selected studies. The experimental designs partly contributed to the observed variation. Therefore, recombinant vaccines that use new carrier system technologies and delivered through oral route in feeds would have been of great value for use in the prevention and control of A. hydrophila infections in fish. Despite the usefulness as academic tools to identify what is important in pathogenicity of the etiological agent to the host fish, these vaccines are only economically viable in very high-value animals. Therefore, if vaccination is a good option for A. hydrophila group, then simple autogenous vaccines based on accurate typing and evidence-based definition of the epidemiological unit for their use would be the most viable approach in terms of both efficacy and economic feasibility especially in low and middle-income countries (LMIC).

A novel rapid direct haemagglutination-inhibition assay for measurements of humoral immune response against non-haemagglutinating Fowlpox virus strains in vaccinated chickens.

Fowlpox (FP) is a serious disease in chickens caused by Fowlpox virus (FPV). One method currently used to control FPV is vaccination followed by confirmation that antibody titres are protective using the indirect haemagglutination assay (IHA). The direct haemagglutination inhibition (HI) assay is not done because most FPV strains do not agglutinate chicken red blood cells (RBCs). A novel FPV strain TPV-1 which agglutinates chicken RBCs was discovered recently and enabled a direct HI assay to be conducted using homologous sera. This study is therefore aimed at assessing the direct HI assay using a recently discovered novel haemagglutinating FPV strain TPV-1 in chickens vaccinated with a commercial vaccine containing a non-haemagglutinating FPV.Chicks vaccinated with FPV at 1 day-old had antibody geometric mean titres (GMT) of log2 3.7 at 7 days after vaccination and log2 8.0 at 28 days after vaccination when tested in the direct HI. Chickens vaccinated at 6 weeks-old had antibody geometric mean titres (GMT) of log2 5.0 at 7 days after vaccination and log2 8.4 at 28 days after vaccination when tested in the direct HI. The GMT recorded 28 days after vaccination was slightly higher in chickens vaccinated at 6-week-old than in chicks vaccinated at one-day-old. However, this difference was not significant (P > 0.05). All vaccinated chickens showed "takes". No antibody response to FPV and "takes" were detected in unvaccinated chickens (GMT < 1). There was a slightly higher GMT in chickens of all ages throughout the observation period when the standard assay, the passive (indirect) haemagglutination was used (Overall GMT reached log2 9.3 ±.0.3 on day 28). However, the difference between the two assays was not significant (P > 0.05). CONCLUSION: These findings indicate that a simple and rapid direct HI assay using the FPV TPV-1 strain as antigen may be used to measure antibody levels in chickens vaccinated with non-haemagglutinating strains of FPV, and that the titres are comparable to those obtained by indirect IHA.

Existence of variant strains Fowlpox virus integrated with Reticuloendotheliosis virus in its genome in field isolates in Tanzania.

Fowlpox virus (FPV) is one example of poultry viruses which undergoes recombination with Reticuloendotheliosis virus (REV). Trepidation had been raised, and it was well established on augmented pathogenicity of the FPV upon integration of the full intact REV. In this study, we therefore intended at assessing the integration of REV into FPV genome of the field isolates obtained in samples collected from different regions of Tanzania. DNA extraction of 85 samples (scabs) was performed, and FPV-specific PCR was done by the amplification of the highly conserved P4b gene. Evaluation of FPV-REV recombination was done to FPV-specific PCR positively identified samples by amplifying the env gene and REV long terminal repeats (5' LTR). A 578-bp PCR product was amplified from 43 samples. We are reporting for the first time in Tanzania the existence of variant stains of FPV integrated with REV in its genome as 65 % of FPV identified isolates were having full intact REV integration, 21 % had partial FPV-REV env gene integration and 5 % had partial 5' LTR integration. Despite of the fact that FPV-REV integrated stains prevailed, FPV-REV-free isolates (9 %) also existed. In view of the fact that full intact REV integration is connected with increased pathogenicity of FPV, its existence in the FPV genome of most field isolates could have played a role in increased endemic, sporadic and recurring outbreaks in selected areas in Tanzania.

Sanitary practices and occurrence of zoonotic conditions in cattle at slaughter in Morogoro Municipality, Tanzania: implications for public health.

As meat consumption is increasing worldwide to cover for protein demands, also raise concerns and challenges regarding meat hygiene and safety. The current one year follow up study aimed at investigating on sanitary practices and occurrence of zoonotic conditions, during post-mortem examination, in cattle at slaughter in Morogoro Municipality abattoir. Sanitary practices were assessed through direct observation where as routine post-mortem inspection procedures were employed to detect zoonotic conditions in cattle at slaughter. During the study period a total of 30,713 cattle were slaughtered and inspected at the abattoir. Results revealed poor hygienic practices at the level of abattoir surrounding, the slaughter operation area, personnel as well as meat vans. Whole carcasses, lungs, livers, hearts and heads were condemned due to zoonotic conditions at rates of 0.026%, 1.96%, 1.61%, 0.02% and 0.21% respectively. Bovine tuberculosis, Cysticercus bovis cysts, fasciolosis and hydatidosis were the responsible zoonotic conditions for the condemnations. Bovine tuberculosis was a leading zoonotic cause of condemnations accounting for 95.7% of lungs and 100% of all head and carcass condemnations. Cysticercus bovis cysts were ranking the second in serving as causes of condemnations closely followed by fasciolosis and lastly hydatidosis. Occurrence of disease conditions with zoonotic implication in cattle at slaughter, meant for human consumption, may pose significant economic and public health risks to especially disaster-prone marginalized communities. Thus, there is a need to introduce appropriate control measures of livestock diseases to minimize the rate of infection; and eventually reduce economic losses and safeguard public health.

Sanitary Practices and Occurrence of Zoonotic Conditions in Cattle at Slaughter in Morogoro Municipality; Tanzania: Implications for Public Health

Abstract:As meat consumption is increasing worldwide to cover for protein demands; also raise concerns and challenges regarding meat hygiene and safety. The current one year follow up study aimed at investigating on sanitary practices and occurrence of zoonotic conditions; during postmortem examination; in cattle at slaughter in Morogoro Municipality abattoir. Sanitary practices were assessed through direct observation where as routine post-mortem inspection procedures were employed to detect zoonotic conditions in cattle at slaughter. During the study period a total of 30;713 cattle were slaughtered and inspected at the abattoir. Results revealed poor hygienic practices at the level of abattoir surrounding; the slaughter operation area; personnel as well as meat vans. Whole carcasses; lungs; livers; hearts and heads were condemned due to zoonotic conditions at rates of 0.026; 1.96; 1.61; 0.02 and 0.21 respectively. Bovine tuberculosis; Cysticercus bovis check for this species in other resources cysts; fasciolosis and hydatidosis were the responsible zoonotic conditions for the condemnations. Bovine tuberculosis was a leading zoonotic cause of condemnations accounting for 95.7 of lungs and 100 of all head and carcass condemnations. Cysticercus bovis cysts were ranking the second in serving as causes of condemnations closely followed by fasciolosis and lastly hydatidosis. Occurrence of disease conditions with zoonotic implication in cattle at slaughter; meant for human consumption; may pose significant economic and public health risks to especially disaster-prone marginalized communities. Thus; there is a need to introduce appropriate control measures of livestock diseases to minimize the rate of infection; and eventually reduce economic losses and safeguard public health