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Queensley Eghianruwa,1,2 Omolaja Osoniyi,3 Naomi Maina,1,4 Sabina Wachira,5 Mabel Imbuga4 1Department of Molecular Biology and Biotechnology, Pan African University Institute of Science, Technology and Innovation, JKUAT campus, Juja, Kenya; 2Department of Biochemistry, University of Uyo, Uyo, Nigeria; 3Department of Biochemistry and Molecular Biology, Obafemi Awolowo University, Ile Ife, Nigeria; 4Biochemistry Department, Jomo Kenyatta University of Agriculture and Technology, Juja, Kenya; 5Centre for Traditional Medicine and Drug Research, Kenya Medical Research Institute, Nairobi, KenyaCorrespondence: Queensley EghianruwaDepartment of Biochemistry, University of Uyo, Uyo, NigeriaTel +2348024195501Email qaeghianruwa@uniuyo.edu.ngPurpose and Methods: In this study, the analgesic activity of the crude alcohol (acetone-methanol) and aqueous (in PBS, pH 7.2) extracts of the marine molluscs, Pachymelania aurita and Tympanotonus fuscatus, has been evaluated using the formalin test (for chronic antinociceptive) and the tail-flick (acute antinociceptive) pain models in male swiss albino mice.Results: The results show that the extracts of P. aurita and T. fuscatus demonstrated high safety margins as single doses of up to 2000 mg/kg bwt proved to be well tolerated and non-lethal, although the alcohol extract of P. aurita caused necrosis in the liver and kidney when administered at a dose level of 2000 mg/kg bwt. In the formalin test, treatment with the aqueous extracts of P. aurita and T. fuscatus as well as the alcohol extract of T. fuscatus 30 min before the subcutaneous injection of 5% formalin to the paw of the mice resulted in a significant time- and dose-dependent reduction in total and phase 2a pain-related behavior and thus nociception. The extracts had no analgesic effect in tail-flick test up to the highest dose tested.Conclusion: Hence, the results from both models indicate that the site of their analgesic action is probably peripheral.Keywords: Nigerian periwinkle, formalin test, anti-nociception and analgesia, marine natural products, bioactivity

Abstract Background Literature is scarce on the occurrence of bovine mastitis and antimicrobial resistance among dairy animals kept by pastoralists in the Kenya. Objectives A cross‐sectional study was carried out to investigate the prevalence and risk factors of subclinical mastitis (SCM) and evaluate the antibiotic sensitivity of bacteria isolated from dairy cattle kept by farmers in Kajiado Central sub‐county, Kenya. Methods A total of 202 lactating cows from 40 farms were sampled. Milk from the cows was screened for SCM using the California mastitis test, and the bacteria present in the milk samples were determined using standard bacteriological methods. The sensitivity of the isolated coagulase‐negative staphylococci (CNS) and Staphylococcus aureus against antibiotics was tested using the Kirby–Bauer disk diffusion method. Results The prevalence of SCM at quarter‐ and cow‐level was 31.7% and 53%, respectively. The prevalence of SCM was significantly higher (p

Background and Aim: The emergence of drug-resistant strains of Eimeria spp. calls for the development of novel anticoccidial drugs. Plant extracts provide a possible natural source for such drugs. This study aimed to investigate the in vitro anticoccidial activity of encapsulated bromelain (EB) in chitosan nanocarriers on Eimeria spp. oocysts isolated from goats kept by farmers in Kenya. Materials and Methods: Bromelain was extracted from the peel of ripe pineapples using standard methods. Eimeria spp. oocysts were isolated from the feces of goats using a flotation method. The inhibition of sporulation was assayed after exposing the oocysts to solutions of EB, non-EB (NEB), and diclazuril (positive control) at concentrations between 4 mg/mL and 0.125 mg/mL for 48 h. The oocysts were examined under a microscope (40x) to determine the effects of the drugs on the sporulation process. The percentage of sporulation inhibition was calculated after 48 h and the inhibition concentration 50% (IC50) was determined by probit analysis. Results: Bromelain manifested anticoccidial activity through the inhibition of the sporulation of coccidia oocysts. EB achieved inhibition with a lower dose compared with NEB. The IC50 values of diclazuril, EB, and NEB were 0.078 mg/mL, 0.225 mg/mL, and 0.575 mg/mL, respectively. There were significant differences (p

This study investigated fluctuations in hematological values of 50 wild-caught vervet monkeys (African green monkeys, grivets, Chlorocebus aethiops) during habituation to captivity. The monkeys were categorized into four groups according to age and sex viz adult males, adult females, juvenile males, and juvenile females. The erythrocyte values were significantly higher (P<0.05) in the adult males than in the other animals. There was an increase in most of the erythrocyte parameters studied during the monitoring period with the most significant being hemoglobin, hematocrit, and mean corpuscular volume. However, the red cell distribution widths, which were higher in adult females, declined. The total white blood cell (WBC) counts, which were higher in adult females than in the other animals, were closely correlated with granulocytes counts. The WBC levels decreased in all the animals throughout the 8 months study, indicating gradually decreasing stress, but they were relatively stable in males. The platelet counts declined significantly (P<0.05) and at 8 months post capture the counts were higher in females than in males. The juvenile female platelet counts were relatively stable during the monitoring period. The maintenance of the monkeys on an improved stable diet and in environment-controlled housing combined with progressing psycho-physiological adaptation may be important factors for the gradual improvements of the hematological values recorded. There were wide variations in these between individual animals emphasizing the need for long adaptation combined with establishment of individual baseline values before experimental studies. Am. J. Primatol. 69:1053–1063, 2007. © 2007 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

Background: A key objective in basic research on human African trypanosomiasis (HAT) is developing a cheap and reliable experimental model of the disease for use in pathogenesis and drug studies. Objective: With a view to improving current models, a study was undertaken to characterise the virulence and pathogenicity of three Trypanosoma brucei rhodesiense stabilates, labelled as International Livestock Research Institute (ILRI)-2918, ILRI-3953, and Institute of Primate Research (IPR)-001, infected into Swiss white mice. Methods: Swiss white mice were infected intraperitoneally with trypanosomes and observedfor parasitaemia using wet blood smears obtained by tail snipping. Induction of late-stagedisease was undertaken using diminazene aceturate (40 mg/kg, Berenil) with curativetreatment done using melarsoprol (3.6 mg/kg, Arsobal). Results: The prepatent period for the stabilates ranged from three to four days with mean peak parasitaemia ranging from Log10 6.40 to 8.36. First peak parasitaemia for all stabilates varied between six and seven days post infection (DPI) followed by secondary latency in ILRI-2918 (15–17 DPI) and IPR-001 (17–19 DPI). Survival times ranged from six DPI (ILRI-3953) to 86 DPI (IPR-001). Hindleg paresis was observed in both ILRI-3953 (at peak parasitaemia) and ILRI-2918 (after relapse parasitaemia). Mice infected with IPR-001 survived until 54 DPI when curative treatment was undertaken. Conclusions: This study demonstrated that the stabilates ILRI-2918 and ILRI-3953 were unsuitable for modelling late-stage HAT in mice. The stabilate IPR-001 demonstrated the potential to induce chronic trypanosomiasis in Swiss white mice for use in development of a late-stage model of HAT.

Background: There are three subspecies of Trypanosoma brucei: T. b. gambiense, T. b. rhodesiense and T. b. brucei. The first two are infectious to humans, whilst T. b. brucei is not. Identifying an animal model of T. b. brucei that mimics human African trypanosomiasis (HAT) would enable researchers to study HAT without subjecting themselves to undue risks such as accidental infection. Objectives: This study assessed the sequential clinical, parasitological and haematological changes in vervet monkeys infected with T. b. brucei. Methods: Three vervet monkeys were infected with a 104 inoculum of T. b. brucei (isolate GUTat 1). Late-stage disease was induced by subcurative treatment with diminazene aceturate 28 days post-infection. The animals were treated curatively with melarsoprol upon relapse. Parasitaemia and clinical signs were monitored daily and, at weekly intervals, the monkeys’ blood and cerebrospinal fluid (CSF) were sampled for haematology and parasitosis assessments, respectively. Results: The first-peak parasitaemia was observed between seven and nine days post-infection. Clinical signs associated with the disease included fever, dullness, pallor of mucous membranes, lymphadenopathy, splenomegaly and oedema. Late-stage signs included stiffness of joints and lethargy. The monkeys developed a disease associated with microcytic hypochromic anaemia. There was an initial decline, followed by an increase, in total white blood cell counts from early- to late-stage disease. Trypanosomes were detected in the CSF and there was a significant increase in white cell counts in the CSF during late-stage disease. Infected vervet monkeys displayed classical clinical symptoms, parasitological and haematological trends that were similar to monkeys infected withT.b. rhodesiense. Conclusion: The T. b. brucei vervet monkey model can be used for studying HAT without putting laboratory technicians and researchers at high risk of accidental infection.

The management of human African trypanosomiasis (HAT) is constrained by lack of simple-to-use diagnostic, staging, and treatment tools. The search for novel biomarkers is, therefore, essential in the fight against HAT. The current study aimed at investigating the potential of IL-6 as an adjunct parameter for HAT stage determination in vervet monkey model. Four adult vervet monkeys (Chlorocebus aethiops) were experimentally infected with Trypanosoma brucei rhodesiense and treated subcuratively at 28 days after infection (dpi) to induce late stage disease. Three noninfected monkeys formed the control group. Cerebrospinal fluid (CSF) and blood samples were obtained at weekly intervals and assessed for various biological parameters. A typical HAT-like infection was observed. The late stage was characterized by significant (P

Helicobacter pylori (H. pylori) strain is the most genetically diverse pathogenic bacterium and now alarming serious human health concern ranging from chronic gastritis to gastric cancer and human death all over the world. Currently, the majority of commercially available diagnostic assays for H. pylori is a challenging task due to the heterogeneity of virulence factors in various geographical regions. In this concern, designing of universal multi-epitope immunogenic biomarker targeted for all H. pylori strains would be crucial to successfully immunodiagnosis assay and vaccine development for H. pylori infection. Hence, the present study aimed to explore the potential immunogenic epitopes of PSA D15 and Cag11 proteins of H. pylori, using immunoinformatics web tools in order to design novel immune-reactive multi-epitope antigens for enhanced immunodiagnosis in humans. Through an in silico immunoinformatics approach, high-ranked B-cell, MHC-I, and MHC-II epitopes of PSA D15 and Cag11 proteins were predicted, screened, and selected. Subsequently, a novel multi-epitope PSA D15 and Cag11 antigens were designed by fused the high-ranked B-cell, MHC-I, and MHC-II epitopes and 50S ribosomal protein L7/L12 adjuvant using linkers. The antigenicity, solubility, physicochemical properties, secondary and tertiary structures, 3D model refinement, and validations were carried. Furthermore, the designed multi-epitope antigens were subjected to codon adaptation and in silico cloning, immune response simulation, and molecular docking with receptor molecules. A novel, stable multi-epitope PSA D15 and Cag11 H. pylori antigens were developed and immune simulation of the designed antigens showed desirable levels of immunological response. Molecular docking of designed antigens with immune receptors (B-cell, MHC-I, MHC-II, and TLR-2/4) revealed robust interactions and stable binding affinity to the receptors. The codon optimized and in silico cloned showed that the designed antigens were successfully expressed (CAI value of 0.95 for PSA D15 and 1.0 for Cag11) after inserted into pET-32ba (+) plasmid of the E. coli K12 strain. In conclusion, this study revealed that the designed multi-epitope antigens have a huge immunological potential candidate biomarker and useful in developing immunodiagnostic assays and vaccines for H. pylori infection., (© 2024 John Wiley & Sons Ltd.)

Background: Literature is scarce on the occurrence of bovine mastitis and antimicrobial resistance among dairy animals kept by pastoralists in the Kenya., Objectives: A cross-sectional study was carried out to investigate the prevalence and risk factors of subclinical mastitis (SCM) and evaluate the antibiotic sensitivity of bacteria isolated from dairy cattle kept by farmers in Kajiado Central sub-county, Kenya., Methods: A total of 202 lactating cows from 40 farms were sampled. Milk from the cows was screened for SCM using the California mastitis test, and the bacteria present in the milk samples were determined using standard bacteriological methods. The sensitivity of the isolated coagulase-negative staphylococci (CNS) and Staphylococcus aureus against antibiotics was tested using the Kirby-Bauer disk diffusion method., Results: The prevalence of SCM at quarter- and cow-level was 31.7% and 53%, respectively. The prevalence of SCM was significantly higher (p < 0.05) in exotic breeds of cattle and those kept under an extensive system of production. A total of 19 bacterial species were isolated with the majority being CNS (40.1%), S. aureus (15.8%) and Micrococcus spp. (10.4%). S. aureus isolates showed varied resistance to the tested antibiotics with the highest resistance being against ceftazidime (75%), amoxycillin (50%) and streptomycin (46.9%). Several S. aureus isolates were resistant to oxacillin (34.4%) and cefoxitin (12.5%). CNSs were more resistant against ceftazidime (79.1%), amoxycillin (34.6%) and oxacillin (32.1%). Majority (92%-100%) of the Staphylococcus spp. were highly sensitive to ciprofloxacin a fluoroquinolone and augmentin., Conclusions: The high prevalence of SCM and bacteria resistant to antibiotics shows a need for animal health professionals and farmers to develop strategies for the management of mastitis and antibiotic resistance in dairy cows in the study area., (© 2023 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Background: Multiplicity of infection (MOI) is an important measure of Plasmodium falciparum diversity, usually derived from the highly polymorphic genes, such as msp1, msp2 and glurp as well as microsatellites. Conventional methods of deriving MOI lack fine resolution needed to discriminate minor clones. This study used amplicon sequencing (AmpliSeq) of P. falciparum msp1 (Pfmsp1) to measure spatial and temporal genetic diversity of P. falciparum., Methods: 264 P. falciparum positive blood samples collected from areas of differing malaria endemicities between 2010 and 2019 were used. Pfmsp1 gene was amplified and amplicon libraries sequenced on Illumina MiSeq. Sequences were aligned against a reference sequence (NC&#95;004330.2) and clustered to detect fragment length polymorphism and amino acid variations., Results: Children < 5 years had higher parasitaemia (median = 23.5 ± 5 SD, p = 0.03) than the > 5-14 (= 25.3 ± 5 SD), and those > 15 (= 25.1 ± 6 SD). Of the alleles detected, 553 (54.5%) were K1, 250 (24.7%) MAD20 and 211 (20.8%) RO33 that grouped into 19 K1 allelic families (108-270 bp), 14 MAD20 (108-216 bp) and one RO33 (153 bp). AmpliSeq revealed nucleotide polymorphisms in alleles that had similar sizes, thus increasing the K1 to 104, 58 for MAD20 and 14 for RO33. By AmpliSeq, the mean MOI was 4.8 (± 0.78, 95% CI) for the malaria endemic Lake Victoria region, 4.4 (± 1.03, 95% CI) for the epidemic prone Kisii Highland and 3.4 (± 0.62, 95% CI) for the seasonal malaria Semi-Arid region. MOI decreased with age: 4.5 (± 0.76, 95% CI) for children < 5 years, compared to 3.9 (± 0.70, 95% CI) for ages 5 to 14 and 2.7 (± 0.90, 95% CI) for those > 15. Females' MOI (4.2 ± 0.66, 95% CI) was not different from males 4.0 (± 0.61, 95% CI). In all regions, the number of alleles were high in the 2014-2015 period, more so in the Lake Victoria and the seasonal transmission arid regions., Conclusion: These findings highlight the added advantages of AmpliSeq in haplotype discrimination and the associated improvement in unravelling complexity of P. falciparum population structure., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Introduction: The Uganda Ministry of Health recommends facility- and community-based differentiated antiretroviral therapy (DART) models to support person-centred care for eligible clients receiving antiretroviral therapy (ART). Healthcare workers assess client eligibility for one of six DART models upon initial enrolment; however, client circumstances evolve, and their preferences are not routinely adjusted. We developed a tool to understand the proportion of clients accessing preferred DART models and compared the outcomes of clients accessing preferred DART models to the outcomes of clients not receiving preferred DART models., Methods: We conducted a cross-sectional study. A sample of 6376 clients was selected from 113 referrals, general hospitals and health centres purposely selected from 74 districts. Clients receiving ART accessing care from the sampled sites were eligible for inclusion. Healthcare workers interviewed clients (caretakers of clients under 18), over a 2-week period between January and February 2022 using a client preference tool to elicit whether clients were receiving DART services through their preferred model. Treatment outcomes of viral load test, viral load suppression and missed appointment date were extracted from clients' medical files before or immediately after the interview and de-identified. The descriptive analysis determined the interaction between client preferences and predefined treatment outcomes by comparing outcomes of clients whose care aligned with their preferences to outcomes of clients whose care misaligned with their preferences., Results: Of 25% (1573/6376) of clients not accessing their preferred DART model, 56% were on facility-based individual management and 35% preferred fast-track drug refills model. Viral load coverage was 87% for clients accessing preferred DART models compared to 68% among clients not accessing their preferred model. Viral load suppression was higher among clients who accessed the preferred DART model (85%) compared to (68%) clients who did not access their preferred DART model. Missed appointments were lower at 29% for clients who accessed preferred DART models compared to 40% among clients not enrolled in the DART model of their choice., Conclusions: Clients who accessed their preferred DART model have better clinical outcomes. Preferences should be integrated throughout health systems, improvement interventions, policies and research efforts to ensure client-centred care and client autonomy., (© 2023 The Authors. Journal of the International AIDS Society published by John Wiley & Sons Ltd on behalf of the International AIDS Society.)

Globally, the emergence of the coronavirus disease (COVID-19) has had a significant impact on life. The need for ongoing SARS-CoV-2 screening employing inexpensive and quick diagnostic approaches is undeniable, given the ongoing pandemic and variations in vaccine administration in resource-constrained regions. This study presents results as proof of concept to use hybridization chain reaction (HCR) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a complex for detecting SARS-CoV-2. HCR hairpin probes were designed using the NUPACK web-based program and further used to amplify the SARS-CoV-2 N gene in archived nasopharyngeal samples. The results were visualized using agarose gels and CRISPR Cas12a-based lateral flow strips. The assay was evaluated using the gold standard, real-time polymerase chain reaction (RT-PCR), as recommended by the World Health Organization (WHO). The results show the comparative efficiency of HCR to RT-PCR. This study shows that HCR and CRISPR are viable alternatives for diagnosing SARS-CoV-2 in samples.

Antimicrobial resistance (AMR) is a growing health problem globally. To address this challenge, there is a need to generate baseline data on the prevalence and AMR profile of the main disease-causing bacteria. Here, we interrogated the prevalence of bacteria in the nasal cavity of healthy pastoralists in Kajiado Central Subcounty, Kenya, and the occurrence of AMR in Staphylococcus isolates among the study subjects. Nasal swabs from 176 pastoralists were cultured, and the bacteria isolates identified using standard phenotypic and biochemical bacteriological methods. Among the obtained 195 isolates, the most prevalent isolates were coagulase-negative Staphylococcus (CoNS) (44.9%), followed by Enterococci spp. (43.2%) while Staphylococcus aureus prevalence was 8%. Antimicrobial sensitivity of the Staphylococcus spp. isolates to 14 antibiotics representing six antibiotic groups was undertaken using the Kirby-Bauer disk diffusion method. Among the CoNS, the highest resistance was reported in amoxicillin (78.7%) and ceftazidime (76%), while the most resistance for S. aureus was reported in ceftazidime (100%), amoxicillin (71.4%), and streptomycin (71.4%). From an administered questionnaire looking at gender, animal contact frequency, history of hospital visitation and antibiotic usage, and habitual intake of raw milk, the study showed that male participants had a higher risk of carrying multiple drug resistant (MDR) bacteria than females ( p = 0.02, OR = 1.3). Likewise, habitual intake of raw milk was significantly associated MDR acquisition ( p = 0.02, OR = 1.82). This study reveals a high prevalence of AMR Staphylococcus isolates in the study area laying a foundation for further analysis of molecular characterization of the observed resistance as well as the development of interventions that can reduce the occurrence of AMR in the study area., Competing Interests: The authors declare that there is no conflict of interest., (Copyright © 2023 Edidah Ong'era et al.)

Dairy ruminant milk provides a conducive environment for bacterial proliferation. In animals, these bacteria are exposed to antibiotics, whose overuse has led to increased cases of drug resistance. A cross-sectional study was conducted on milk and milk products vended in Juja Sub-County, Kenya to determine the prevalence of bacteria and antibiogram of Staphylococcus spp. and Escherichia coli . A total of 169 milk samples were obtained from various outlets in the study area. Milk samples were cultured and isolated bacteria were identified using standard bacteriological procedures. Various bacteria (15 species) were isolated in different proportions. Staphylococcus spp. and E. coli were isolated from 25.4% and 11.8% of the collected samples, respectively. The highest number of Staphylococcus spp. were isolated from raw milk ( n  = 34) while the highest number of E. coli where isolated from fermented milk ( n  = 15). Staphylococcus spp. and E. coli isolates were subjected to antimicrobial susceptibility tests using CLSI guidelines. The Staphylococcus spp. isolates were highly resistant to penicillin G (93%) but susceptible to norfloxacin (100%), gentamicin (90.6%), and chloramphenicol (86%). The E. coli isolates were highly resistant to cephalexin (85%) and ceftazidime (60%) but susceptible to chloramphenicol (100%), norfloxacin (95%), gentamicin (95%), azithromycin (95%) and cefepime (80%). Furthermore, 44.3% of Staphylococcus spp. and 50% of E. coli isolates had a Multiple Antibiotic Resistance (MAR) Index greater than 0.2. This implies that these bacteria were high-risk bacteria whose treatment with current antibiotics would be challenging. The high prevalence and multidrug resistance patterns shown by the Staphylococcus spp. and E. coli isolated from milk products in Juja Sub-county highlights the importance of proper handling and processing of milk from the farm to consumers. This will in turn reduce the possibility of zoonotic transfer of multidrug-resistant bacteria., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Julius I. N. Sombie et al.)

Objective: This cross-sectional study investigated the circulating strains of rotavirus and screened for noravirus in Ibadan, Nigeria as the country introduces the rotavirus vaccine into its national immunization program., Methods: Sixty-five stool samples were collected from children younger than 5 years with clinically diagnosed diarrhea and screened for the presence of rotavirus and norovirus using RT-PCR. Rotavirus-positive samples were further analyzed to determine the G and P genotypes using semi-nested multiplex PCR., Results: The rates of rotavirus and norovirus positivity were 30.8% and 10.8%, respectively, whereas the rate of rotavirus and norovirus mixed infection was 4.6%. G1 was the predominant VP7 genotype, followed by G2, G9, and G1G2G9, whereas the predominant VP4 genotype was P[4], followed by P[6], P[8], and P[9]. The mixed P types P[4]P[8] and P[4]P[6] were also detected. G1P[4] was the most common VP4 and VP7 combination, followed by G2P[4], G1[P6], G1P[8], G2P[6], G2P[9], G9P[6], G2G9P[4], G2P[4]P[6], G1P[4]P[8], G2G9P[8], G1G2G9P[8], and G1[non-typable] P[non-typable], which were detected in at least 5% of the samples. Four samples had a combination of non-typable G and P types., Conclusions: It is essential to monitor the circulation of virus strains prior to and during the implementation of the immunization program.

Background and Aim: The emergence of drug-resistant strains of Eimeria spp. calls for the development of novel anticoccidial drugs. Plant extracts provide a possible natural source for such drugs. This study aimed to investigate the in vitro anticoccidial activity of encapsulated bromelain (EB) in chitosan nanocarriers on Eimeria spp. oocysts isolated from goats kept by farmers in Kenya., Materials and Methods: Bromelain was extracted from the peel of ripe pineapples using standard methods. Eimeria spp. oocysts were isolated from the feces of goats using a flotation method. The inhibition of sporulation was assayed after exposing the oocysts to solutions of EB, non-EB (NEB), and diclazuril (positive control) at concentrations between 4 mg/mL and 0.125 mg/mL for 48 h. The oocysts were examined under a microscope (40x) to determine the effects of the drugs on the sporulation process. The percentage of sporulation inhibition was calculated after 48 h and the inhibition concentration 50% (IC 50 ) was determined by probit analysis., Results: Bromelain manifested anticoccidial activity through the inhibition of the sporulation of coccidia oocysts. EB achieved inhibition with a lower dose compared with NEB. The IC 50 values of diclazuril, EB, and NEB were 0.078 mg/mL, 0.225 mg/mL, and 0.575 mg/mL, respectively. There were significant differences (p<0.01) between the IC 50 of EB and NEB compared with the standard treatment drug., Conclusion: This preliminary study showed that EB has anticoccidial activity supporting further evaluation at an in vivo level to develop a novel drug for the management of coccidiosis in goats., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Daiba, et al.)

Background: Hereditary angioedema (HAE), a rare disease that is characterized by painful and recurring non-allergic swelling episodes, is caused by the deficiency or dysfunction of C1 inhibitor (C1INH) protein. A comprehensive HAE management plan may require long-term prophylaxis (LTP) in addition to on-demand treatment to help "normalize" patients' lives so that they may fully engage in work, school, family, and leisure activities., Aim: The main objective of this narrative review is to provide an overview of updated guideline recommendations specific to LTP of HAE and discuss clinical considerations and pharmacologic management options, with a focus on C1INH., Materials and Methods: The authors reviewed relevant HAE literature for current recommendations regarding LTP and the role of C1NH., Results: Acute HAE attacks are treated with on-demand medication; however, there is a consensus that LTP should routinely be considered for risk reduction and prevention of future episodes. The 2017 World Allergy Organization/European Academy of Allergy and Clinical Immunology guidelines recommend that all patients with HAE be evaluated for LTP routinely and the 2020 HAE Association (HAEA) guidelines emphasize that the decision to use LTP should not be based on rigid criteria, but rather should be based on individual patient needs. Both guidelines recommend C1INH as first-line/preferred therapy for LTP in a range of patient types including adults, children/adolescents, and pregnant/lactating patients. The HAEA also recommends the kallikrein inhibitor, lanadelumab, as a first-line option for LTP. HAE pathway-specific agents for LTP have not been associated with notable safety concerns., Discussion: Plasma-derived C1INH has been available for 40+ years in Europe and impacts multiple targets within the HAE pathway. C1INH has been used for on-demand treatment and LTP. A subcutaneous formulation of plasma-derived C1INH is approved for LTP and produces functional C1INH activity levels consistently above the threshold needed for protection from HAE attacks. Other pathway-specific options for LTP include the plasma kallikrein inhibitors, lanadelumab-flyo and berotralstat, approved for adults and pediatric patients aged ≥12 years. C1INH is approved for adults and pediatric patients aged ≥6 years., Conclusion: Assessing the need for LTP is vital in the ongoing dialogue between clinicians and patients, as both disease-related factors and patient preferences may change over time. Among available options for LTP, plasma-derived C1INH is the broadly recommended first-line option for LTP in patients with HAE, including pregnant/lactating women and pediatric patients (≥6 years)., Competing Interests: John Anderson has served as a speaker for, has been on advisory boards for, and has worked on clinical research with Pharming, CSL Behring, Shire/Takeda, and BioCryst; he has been on an advisory board for BioMarin. Njeri Maina reports no conflicts of interest., (© 2022 The Authors. Clinical and Translational Allergy published by John Wiley & Sons Ltd on behalf of European Academy of Allergy and Clinical Immunology.)

Zoonotic pathogens are among the most important causes of ill health all over the world. The presence of these pathogens in free ranging baboons may have significant implications for humans. In Kenya, the encroachment of wildlife habitats has led to increased interaction between humans and wildlife especially non-human primates. The current study therefore aimed at investigating any possible zoonotic gastrointestinal helminths of olive baboons (Papio anubis) at the human-wildlife interface in two park borders and a ranch in Kenya, namely, Tsavo West National Park, Tana River Primate Reserve and Mutara Ranch, Laikipia, Kenya. One hundred and forty-seven baboons were used in the study. They were trapped in the wild, sampled for stool marked and then released back to the wild. Gastrointestinal (GIT) helminths identified were Strongyloides, Oesophagostomum, Enterobius spp and Trichuris Trichiura from all the three sites while Schistosoma mansoni was only detected from Tsavo baboons and with very low incidence (2.1%). The prevalence of these parasites varied among the sites but significant difference in prevalence was only noted in Strongyloides and Oesophagostomum (p < 0.05) among the three sites. This therefore implies that even with control measures instituted on the human population, baboons will always be a source of zoonotic GIT helminths especially S. mansoni even if the incidence are low. There is need to put in place measures aiming to reduce their interactions with humans and also try to control these infections in the baboons., (© 2021 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Wood decomposition is a sophisticated process where various biocatalysts act simultaneously and synergistically on biopolymers to efficiently break down plant cell walls. In nature, this process depends on the activities of the wood-inhabiting fungal communities that co-exist and interact during wood decay. Wood-decaying fungal species have traditionally been classified as white-rot and brown-rot fungi, which differ in their decay mechanism and enzyme repertoire. To mimic the species interaction during wood decomposition, we have cultivated the white-rot fungus, Bjerkandera adusta , and two brown-rot fungi, Gloeophyllum sepiarium and Antrodia sinuosa , in single and co-cultivations on softwood and hardwood. We compared their extracellular hydrolytic carbohydrate-active and oxidative lignin-degrading enzyme activities and production profiles. The interaction of white-rot and brown-rot species showed enhanced (hemi)cellulase activities on birch and spruce-supplemented cultivations. Based on the enzyme activity profiles, the combination of B. adusta and G. sepiarium facilitated birch wood degradation, whereas B. adusta and A. sinuosa is a promising combination for efficient degradation of spruce wood, showing synergy in β-glucosidase (BGL) and α-galactosidase (AGL) activity. Synergistic BGL and AGL activity was also detected on birch during the interaction of brown-rot species. Our findings indicate that fungal interaction on different woody substrates have an impact on both simultaneous and sequential biocatalytic activities.

Hereditary angioedema (HAE) is a rare disease. Regardless, patients with HAE have access to multiple state-of-the-art medications available for on-demand use and prevention that reduce the frequency and burden of HAE attacks. These treatments have greatly reduced the burden of disease and helped patients achieve improved quality of life. However, with greater numbers of therapeutic options, HAE care has become more complex. In this review, we addressed essential elements of an individualized comprehensive management plan for a patient with HAE. We focused on access to an expert physician, ongoing patient education, access to effective treatment options, coordination of care and management of treatment logistics, ongoing monitoring of attacks and treatments, and other resources for patient support. This plan will need to be communicated with the patient and other care providers, especially during emergent conditions, and accommodate the patient's lifestyle with consideration for work, school, travel, etc. Periodically, the physician and the patient will need to review information about attacks, triggers, and treatments to identify areas for improvement and update the plan.

Aim: This study determined the resistance pattern to β-lactam antibiotics of bacteria isolated from goats with subclinical mastitis in Thika subcounty, Kenya. We also administered a questionnaire to assess the risk factors associated with the occurrence of resistance to commonly used antibiotics., Materials and Methods: We collected milk samples from 110 lactating dairy goats in Thika subcounty to screen for subclinical mastitis using the California mastitis test. Bacterial isolation and identification were performed according to colony morphology, the hemolytic pattern on sheep blood agar, lactose fermentation on MacConkey plates, Gram staining, and standard biochemical tests. The antibiotic susceptibility of the isolates was determined by the agar disk diffusion method using penicillin G, cephalexin, cefoxitin, and cefotaxime antibiotic disks. The double-disk synergy test using amoxicillin-clavulanic acid was employed as a confirmatory test for extended-spectrum β-lactamase (ESBL) production. Fisher's exact test was used to determine the risk factors associated with the occurrence of antibiotic resistance (p≤0.05 was considered significant)., Results: Of the 110 dairy goats sampled, 72.7% (80) were positive for subclinical mastitis. Isolation and identification of the bacteria from the positive samples yielded 149 bacteria isolates, including Staphylococcus aureus , Klebsiella pneumoniae , Acinetobacter spp., Yersinia spp., coagulase-negative staphylococci, and Escherichia coli . A high percentage (76.5%, 114/149) of the bacterial isolates was resistant to at least one of the tested antibiotics. At least 56/106 isolates (52.8%) showing cross-resistance to the β-lactam antibiotics were resistant to all four of the tested antibiotics, while only one isolate was resistant to three antibiotics (penicillin G, cephalexin, and cefoxitin). The double-disk synergy test confirmed that none of the isolates possessed ESBLs. Pre- and post-milking practices (p=0.0336) were found to be significantly associated with the occurrence of antibiotic resistance., Conclusion: A large proportion of the goats in our study cohort were infected with β-lactam-resistant bacteria associated with subclinical mastitis. Because the identified bacteria are of zoonotic importance, further studies should be undertaken to determine the transmission dynamics between humans and livestock and to identify novel intervention strategies., (Copyright: © Okoko, et al.)

Background and Aim: The development of resistance to anthelmintic drugs has prompted research into alternative methods of controlling intestinal nematodes in ruminants. This study aimed at evaluating the in vitro and in vivo anthelmintic efficacy and toxicity of chitosan encapsulated bromelain in Small East African goats in Kenya., Materials and Methods: Adult mortality assay was performed using live Haemonchus contortus worms treated with encapsulated bromelain solution ranging from 0.125 mg/ml to 2 mg/ml. Percentage mortality of worms was calculated after 24 h and the lethal concentration 50% (LC 50 ) determined. For the in vivo study, 18 healthy male indigenous goats were divided into six groups of three goats each. The encapsulated bromelain was orally administered in increasing dosages (3-30 mg kg) once daily, for 14 days. The packed cell volume (PCV), aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, creatinine, and fecal egg count (FEC) were determined on a weekly basis. At the end of the study, the goats were sacrificed and gross pathology and histopathology of main organs assessed., Results: Albendazole had the highest (p<0.05) anthelmintic effect on the worms. An LC 50 of 0.05 mg/ml, 0.445 mg/ml, and 0.155 mg/ml was observed for albendazole, plain bromelain, and encapsulated bromelain, respectively. The PCV of treated and untreated goats did not show any significant difference (p>0.05), varied from 29.3% to 35.1%, and was within the normal range of the animal. Likewise, no significant differences (p>0.05) were observed between the AST, ALT, urea, and creatinine levels of treated and the control (non-treated) goats. No adverse clinical symptoms, toxicity of the main organs, and mortality in goats were associated with the chitosan encapsulated bromelain after administration of dose up to 30 mg/kg for 14 days. Therefore, the lethal dose 50 of encapsulated bromelain may be considered to be >30 mg/kg. On day 28 post-treatment, the encapsulated bromelain showed a higher in vivo FEC reduction (68.8%) as compared to the plain bromelain (32.4%)., Conclusion: Our results show that bromelain encapsulated in chitosan may be safe and effective in reducing the burden of gastrointestinal tract strongyle nematodes in goats. However, there is a need for further studies to establish the dosage of the encapsulated bromelain to be administered in a single dose for the treatment of goats against gastrointestinal strongyles. In addition, species-specific studies on the efficacy of encapsulated bromelain on strongyles are necessary to evaluate its effectiveness against the entire Strongyloididae family., (Copyright: © Wasso, et al.)

Enzyme linked immunosorbent assays (ELISAs) have been developed for the detection of antibodies against contagious caprine pleuropneumonia (CCPP), the causative agent of which is Mycoplasma capricolum subsp. Capripneumoniae (Mccp). The currently available commercial CCPP competitive ELISA (CCPP cELISA) kit produced and supplied by IDEXX Company (Westbrook, Maine, United States) is relatively expensive for most African laboratories. To address this issue and provide a variety of choices, a sensitive and specific blocking-ELISA (b-ELISA) test to detect antibodies against CCPP was developed. We describe the newly developed CCPP blocking-ELISA based on the blocking of an epitope of a monoclonal antibody (Mccp-25) by a positive serum sample against the Mccp protein coated on a plate. The Percentage Inhibition (PI) cut-off value for the CCPP b-ELISA was set at 50 using 466 CCPP negative and 84 CCPP positive small ruminant sera. Of the negative sera, 307 were obtained from the Botswana National Veterinary Laboratory (BNVL) and 159 from the Friedrich-Loeffler-Institute (FLI) Germany. The 84 positive sera samples came from experimentally vaccinated goats at the AU-PANVAC facility in Debre-Zeit, Ethiopia. The relative diagnostic sensitivity and specificity of the CCPP b-ELISA was 93% and 88%, respectively. This test result indicated good correlation with that of the commercial CCPP cELISA by IDEXX Company (Westbrook, Maine, United States) with a Cohen's κ agreement of κ agreement of 0.85. The newly developed CCPP b-ELISA will be useful in the detection of antibodies for the diagnosis CCPP and for sero-surveillance during vaccination campaigns.

This study aimed to investigate the antimitotic and antiproliferation activities of crude acetone-methanol and aqueous extracts of two marine molluscs commonly found in the Niger Delta region of Nigeria; T.fuscatus and P.aurita , against human cancerous cell lines (DU145, Hep-2, and HCC1395) cell lines in vitro . The antimitotic activity of the extracts was evaluated using Allium cepa root meristematic cells. Antiproliferative activity of the plant extracts against the cancerous cell lines was compared with normal cell line (VeroE6). Doxorubicin was used as a positive control. Gene expression studies using qPCR for the proapoptotic genes, CASP3, CASP8 and P53 were also carried out. The alcohol extract of T.fuscatus (TFAC) exhibited the most promising activity against all the cancer cell lines tested (DU145 IC 50 = 96.48 ± 1.36 μg/ml, HCC 1395 IC 50 = 61.44 ± 2.45 μg/ml, Hep2 IC 50 = 0.52 ± 0.36 μg/ml) and also had the highest selectivity index of 4.94, 7.78 and 921.97 for DU145, HCC 1395 and Hep-2 cells respectively. Furthermore, TFAC was the only extract that significantly upregulated the expression of caspase 3, caspase 8 and P53. Thus, these findings suggest potential exploitation of TFAC as an anticancer agent., Competing Interests: None.

A cross-sectional study was carried out to determine the prevalence and risk factors of subclinical mastitis in dairy goats in Thika East Subcounty, Kenya. Further the bacterial pathogens and their antibiogram were investigated. Farm level data on risk factors were obtained from 41 farmers using questionnaires. Milk was obtained from 110 lactating dairy goats and tested for submastitis using California Mastitis Test (CMT). The prevalence of subclinical mastitis at goat level was estimated to be at 50.9% using CMT, out of which 86.5% yielded bacteria on culture. The significant risk factors associated with the occurrence of subclinical mastitis were cleaning schedule (p=0.022, OD=1.047) and parity of the goat (p=0048, OD=1.37). Higher prevalence of subclinical mastitis was observed for goats residing in houses cleaned at least once a fortnight. Does in the first parity were least affected. 169 bacterial isolates were obtained from culture, of which 52 isolates from major classes of isolated bacteria were tested for antibiotic sensitivity to six antibiotics. Fourteen different bacteria were isolated and identified from the milk samples. Coagulase-negative Staphylococci (20.7%), Serratia spp. (19.5%), Citrobacter spp. (16%), Klebsiella spp. (11%), Staphylococcus aureus (10.7%), Enterobacter spp. (6.5%), Escherichia coli (5.9%), Proteus spp. (3%), Corynebacterium spp. (1.8%), Morganella spp. (1.8%), Streptococcus spp. (1.2%), Providencia spp. (0.6%), Micrococcus spp. (0.6%), and Staphylococcus intermedius (0.6%) were isolated and identified from the samples. All the isolates were resistant to Penicillin G, while 98% of the isolates were sensitive to Streptomycin. In conclusion, the study showed that a large proportion of goats were affected by subclinical mastitis, with the main bacteria being Staphylococci spp. and coliforms, and that most of the tested antibiotics can be used in the treatment of mastitis. Farmers need to be trained on improved control of mastitis through adoption of good dairy husbandry and milking practices.

Asthma is a chronic inflammatory disease process involving the conductive airways of the human lung. The dysregulated inflammatory response in this disease process may involve multiple cell-cell interactions mediated by signaling molecules, including lipid mediators. Extracellular vesicles (EVs) are lipid membrane particles that are now recognized as critical mediators of cell-cell communication. Here, we compared the lipid composition and presence of specific lipid mediators in airway EVs purified from the bronchoalveolar lavage (BAL) fluid of healthy controls and asthmatic subjects with and without second-hand smoke (SHS) exposure. Airway exosome concentrations were increased in asthmatics, and correlated with blood eosinophilia and serum IgE levels. Frequencies of HLA-DR + and CD54 + exosomes were also significantly higher in asthmatics. Lipidomics analysis revealed that phosphatidylglycerol, ceramide-phosphates, and ceramides were significantly reduced in exosomes from asthmatics compared to the non-exposed control groups. Sphingomyelin 34:1 was more abundant in exosomes of SHS-exposed asthmatics compared to healthy controls. Our results suggest that chronic airway inflammation may be driven by alterations in the composition of lipid mediators within airway EVs of human subjects with asthma.

Infections caused by Mycoplasma species belonging to the 'mycoides cluster' negatively affect the agricultural sector through losses in livestock productivity. These Mycoplasma strains are resistant to many conventional antibiotics due to the total lack of cell wall. Therefore, there is an urgent need to develop new antimicrobial agents from alternative sources such as medicinal plants to curb the resistance threat. Recent studies on extracts from Solanum aculeastrum and Piliostigma thonningii revealed interesting antimycoplasmal activities hence the motivation to investigate the antimycoplasmal activities of constituent compounds. The CH 2 Cl 2 /MeOH extracts from the berries of S. aculeastrum yielded a new β-sitosterol derivative ( 1 ) along with six known ones including; lupeol ( 2 ), two long-chain fatty alcohols namely undecyl alcohol ( 3 ) and lauryl alcohol ( 4 ); two long-chain fatty acids namely; myristic acid ( 5 ) and nervonic acid ( 6 ) as well as a glycosidic steroidal alkaloid; (25R)-3β- O -α-L-rhamnopyranosyl-(1→2)- O -[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranosyloxy-22α-N-spirosol-5-ene ( 7 ) from the MeOH extracts. A new furan diglycoside, (2,5-D-diglucopyranosyloxy-furan) ( 8 ) was also characterized from the CH 2 Cl 2 /MeOH extract of stem bark of P. thonningii . The structures of the compounds were determined on the basis of spectroscopic evidence and comparison with literature data. Compounds 1, 3, 4, 7 , and 8 isolated in sufficient yields were tested against the growth of two Mycoplasma mycoides subsp. mycoides ( Mmm ), two M. mycoides. capri ( Mmc ), and one M. capricolum capricolum ( Mcc ) using broth dilution methods, while the minimum inhibitory concentration (MIC) was determined by serial dilution. The inhibition of Mycoplasma in vitro growth was determined by the use of both flow cytometry (FCM) and color change units (CCU) methods. Compounds 4 and 7 showed moderate activity against the growth of Mmm and Mmc but were inactive against the growth of Mcc . The lowest MIC value was 50 μg/ml for compound 7 against Mmm . The rest of the compounds showed minimal or no activity against the strains of Mycoplasma mycoides tested. This is the first report on the use of combined FCM and CCU to determine inhibition of in vitro growth of Mycoplasma mycoides . The activity of these compounds against other bacterial strains should be tested and their safety profiles determined.