In vitro anticoccidial activity of nanoencapsulated bromelain against Eimeria spp. oocysts isolated from goats in Kenya.

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<0.01) between the IC50 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.

A Review on the Present Advances on Studies of Toxoplasmosis in Eastern Africa.

Toxoplasmosis is a zoonotic infection caused by the protozoan parasite, Toxoplasma gondii. It was discovered over 100 years ago and is credited as the most successful parasitic organism worldwide, able to infect and multiply in all warm blooded animals including an estimated 2.3 billion people. Toxoplasmosis is asymptomatic in immunocompetent individuals. Infection in the developing fetus and immunocompromised individuals can cause severe clinical disease. Toxoplasmosis is also a major cause of reproductive failure in livestock. The economic impact of toxoplasmosis is believed to be substantial. Factors associated with toxoplasmosis infection have been defined. Eastern Africa region is a high-risk area mainly due to the close association of humans and livestock as well as sociocultural practices, poor environmental hygiene, and poverty. The present paper provides a narrative review of published data on toxoplasmosis in Eastern Africa.

Effect of feed supplementation of Dacryodes edulis parts' powder as prebiotic on the growth traits, ceca microbiota and blood parameters of local chickens.

This study aimed at testing the efficacy and safety of Dacryodes edulis plant parts in diets fed to chicken. The plant has potential for use as a natural prebiotic to substitute the conventionally used antibiotic growth promoters in poultry production. Phytochemical analyses of the plant leaves, stem, and bark combination (stembark) and seed powders from the D. edulis were carried out. The powder from the three D. edulis plant parts were used as supplement in formulating six experimental diets tested in this study. The diets were TL0Ed (0.5% leaves powder), TL1Ed (1.0% leaves powder), TB0Ed (0.5% stembark powder), TB1Ed (1.0% stembark powder), TS0Ed (0.5% seeds powder), and TS1Ed (1.0% seeds powder). Besides, a positive (T+ positive control; 0.5-g oxytetracycline as recommended by the manufacturer) and a negative (T- negative control; having no commercial antibiotic and no plant supplement) diets were prepared for comparison purposes. The diets were fed to a total of 288 dual-purpose chicken for a period of 14 weeks. The chicken growth and body composition characteristics, blood chemistry, and microbiota count were collected and used as indicators of the plant parts efficacy and safety. The analysis of the D. edulis plant parts significantly differed (P ≤ 0.05) in their phytochemical contents. The initial body weight and feed conversion efficiency ratios were not significantly different (P ≥ 0.05) between and among treatment groups. However, significant differences (P ≤ 0.05) were detected in the feed intake and body weight gain at eighth week. Live weight at eighth week was significantly different (P ≤ 0.05) with its values ranging between 503.32 and 614.93 g for treatments TL1Ed and TNeg-, respectively. The dietary treatment of D. edulis leaves, stembark, and seed powder at the two inclusion levels significantly (P ≤ 0.05) decreased the colonies forming unit of Escherichia coli and Salmonella sp. as compared with negative control treatment in the eighth week phase. The level of glucose, total cholesterol, triglycerides, aspartate aminotransferase, alanine amino transferase, alkaline phosphatase, and the packed cell volume did not differ significantly (P ≥ 0.05) between and among dietary D. edulis treatments. The findings from this research provide crucial information on the efficacy and safety of D. edulis plant parts. This is an important step in testing the potential of the plant in use as a prebiotic in chicken feeds production.

Prevalence of Toxoplasma gondii and Other Gastrointestinal Parasites in Domestic Cats from Households in Thika Region, Kenya.

Gastrointestinal (GIT) parasites of domestic cats (Felis catus) not only cause morbidity but are also potential zoonotic agents. The current study aimed at establishing the prevalence of GIT parasites in cats kept by households in Thika region, Kenya. Fecal samples were collected randomly from 103 cats and analyzed for presence of parasites using standard parasitological methods. In descending order, the prevalence of the detected protozoa parasites was Isospora spp. 43.7% (95% CI: 40.4-47%), Cryptosporidium spp. 40.8% (95% CI: 37.5-44.1%), Toxoplasma gondii 7.8% (95% CI: 4.5-11.1%), and Entamoeba spp. 2.9% (95% CI: 1.6-6.2%). The prevalence of the observed helminths was Strongyloides stercoralis 43.7% (95% CI: 40.4-47%), Toxocara cati 23.3% (95% CI: 20-26.6%), Ancylostoma spp. 9.7% (95% CI: 6.4-13%), Dipylidium caninum 8.7% (95% CI: 5.4-12.0%), and Acanthocephala spp. 1.9% (95% CI: 1-4.2%). The percentage of cats excreting at least one species of parasite was 73.2% (95% CI = 69.9-76.5%). The study shows that the cats have high spectrum (9) of parasites which are known to affect the cat's health and some are of zoonotic significance.

Development of Neurological Mouse Model for Toxoplasmosis Using Toxoplasma gondii Isolated from Chicken in Kenya.

Animal models for the toxoplasmosis are scarce and have limitations. In this study, a neurological mouse model was developed in BALB/c mice infected intraperitoneally with 15 cysts of a Toxoplasma gondii isolate. The mice were monitored for 42 days and euthanized at different time points. Another group of mice were orally treated with dexamethasone (DXM: 2.66 mg/kg daily, 5.32 mg/kg daily) at 42 days after infection and monitored for a further 42 days. A mortality rate of 15% and 28.6% was observed in mice given 2.66 mg/kg/day and 5.32 mg/kg/day of DXM, respectively. The mean cyst numbers in the brain of DXM treated mice increased up to twofold compared with chronically infected untreated mice. Infections up to 42 days were associated with an increase in both IgM and IgG levels but following dexamethasone treatment, IgM levels declined but IgG levels continued on rising. The brain of toxoplasmosis infected mice showed mononuclear cellular infiltrations, neuronal necrosis, and cuffing. The severity of pathology was higher in mice treated with dexamethasone compared to the positive control groups. The findings of this study demonstrate that DXM-induced reactivation of chronic toxoplasmosis may be a useful development of laboratory animal model in outbred mice used for in vivo studies.

Detection of Natural Toxoplasma gondii Infection in Chicken in Thika Region of Kenya Using Nested Polymerase Chain Reaction.

The detection of Toxoplasma gondii in free-range chickens is a good indicator of possible risk to human beings. The aim of this study was to investigate the occurrence of T. gondii in free-range chicken using polymerase chain reaction (PCR). Brain samples from 105 free-range chickens from three administrative areas in Thika region, Kenya, were collected, DNA-extracted, and analyzed using PCR to detect presence of T. gondii. The overall prevalence of T. gondii in all the three areas was 79.0% (95% CI: 70.0-86.4%) and the prevalence across the three areas was not significantly different (P = 0.5088; χ2 = 1.354). Female chickens had higher (79.4%) prevalence than males (78.6%), although the difference was not significant (P = 0.922, χ2 = 0.01). However, chickens that were more than 2 years old had significantly (P = 0.003; χ2 = 11.87) higher prevalence compared to younger ones. The study indicates that there was a high occurrence of T. gondii infection in free-range chickens from Thika region and that the infection rate is age dependent. Further studies should be carried out to determine the possible role of roaming chickens in the epidemiology of the disease among humans in the area.

Loop Mediated Isothermal Amplification for Detection of Trypanosoma brucei gambiense in Urine and Saliva Samples in Nonhuman Primate Model.

Human African trypanosomiasis (HAT) is a vector-borne parasitic zoonotic disease. The disease caused by Trypanosoma brucei gambiense is the most prevalent in Africa. Early diagnosis is hampered by lack of sensitive diagnostic techniques. This study explored the potential of loop mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) in the detection of T. b. gambiense infection in a vervet monkey HAT model. Six vervet monkeys were experimentally infected with T. b. gambiense IL3253 and monitored for 180 days after infection. Parasitaemia was scored daily. Blood, cerebrospinal fluid (CSF), saliva, and urine samples were collected weekly. PCR and LAMP were performed on serum, CSF, saliva, and urine samples. The detection by LAMP was significantly higher than that of parasitological methods and PCR in all the samples. The performance of LAMP varied between the samples and was better in serum followed by saliva and then urine samples. In the saliva samples, LAMP had 100% detection between 21 and 77 dpi, whereas in urine the detection it was slightly lower, but there was over 80% detection between 28 and 91 dpi. However, LAMP could not detect trypanosomes in either saliva or urine after 140 and 126 dpi, respectively. The findings of this study emphasize the importance of LAMP in diagnosis of HAT using saliva and urine samples.

Comparative pathogenicity of Trypanosoma brucei rhodesiense strains in Swiss white mice and Mastomys natalensis rats.

We evaluated Mastomys natelensis rat as an animal model for Rhodesian sleeping sickness. Parasitaemia, clinical and pathological characteristics induced by T. b. rhodesiense isolates, KETRI 3439, 3622 and 3637 were compared in Mastomys rats and Swiss white mice. Each isolate was intra-peritonially injected in mice and rat groups (n=12) at 1×10(4) trypanosomes/0.2mL. Pre-patent period (PP) range for KETRI 3439 and KETRI 3622-groups was 3-6 days for mice and 4-5 days for rats while for KETRI 3637-infected mice and rats was 5-9 and 4-12 days, respectively. Pairwise comparison between PP of mice and rats separately infected with either isolate showed no significant difference (p>0.05). The PP's of KETRI 3637-infected mice were significantly (p>0.01) longer than those infected with KETRI 3439 or KETRI 3622, a trend also observed in rats. The second parasitaemic wave was more prominent in mice. Clinical signs included body weakness, dyspnoea, peri-orbital oedema and extreme emaciation which were more common in rats. Survival time for KETRI 3439 and 3622-infected groups was significantly (p<0.05) longer in mice than rats but similar in KETRI 3637-infected groups. Inflammatory lesions were more severe in rats than mice. All mice and KETRI 3622-infected rats had splenomegaly, organ congestion with rats additionally showing prominent lymphadenopathy. KETRI 3439-infected rats showed hemorrhagic pneumonia, enteritis with moderate splenomegaly and lymphadenopathy. KETRI 3637-infected rats had the most severe lesions characterized by prominent splenomegaly, lymphadenopathy, hepatomegaly, enlarged adrenal glands, organ congestion, generalized oedemas, gastroenteritis, pneumonia and brain congestion. KETRI 3637-infected Mastomys is a suitable model for studying pathophysiology of HAT.

Erythrina abyssinica prevents meningoencephalitis in chronic Trypanosoma brucei brucei mouse model.

Human African trypanosomiasis is prevalent in Sub-sahara African countries that lie between 14° North and 29° south of the equator. Sixty million people are at risk of infection. Trypanosoma brucei gambesience occurs in West and Central Africa while Trypanosoma brucei rhodesience occurs in East and Southern Africa. The neurological stage of the disease is characterized by neuroinflammation. About 10% of patients treated with the recommended drug, melarsoprol develop post treatment reactive encephalopathy, which is fatal in 50% of these patients, thus melarsoprol is fatal in 5% of all treated patients. This study was aimed at establishing the potential activity of Erythrina abyssinica in reducing neuroinflammation following infection with Trypanosoma brucei brucei. Swiss white mice were divided into ten groups, two control groups and eight infected groups. Infected mice received either methanol or water extract of Erythrina abyssinica at 12.5, 25, 50 or 100 mg/kg body weight. Parasite counts were monitored in peripheral circulation from the third day post infection up to the end of the study. Brains were processed for histology, immunohistochemistry scanning and transmission electron microscopy. Following infection, trypanosomes were observed in circulation 3 days post-infection, with the parasitaemia occurring in waves. In the cerebrum, typical brain pathology of chronic trypanosomiasis was reproduced. This was exhibited as astrocytosis, perivascular cuffing and infiltration of inflammatory cells into the neuropil. However, mice treated with Erythrina abyssinica water extract exhibited significant reduction in perivascular cuffing, lymphocytic infiltration and astrocytosis in the cerebrum. The methanol extract did not have a significant difference compared to the non-treated group. This study provides evidence of anti-inflammatory properties of Erythrina abyssinica and may support its wide use as a medicinal plant by various communities in Kenya.

IL-6 is upregulated in late-stage disease in monkeys experimentally infected with Trypanosoma brucei rhodesiense.

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 < 0.05) elevation of CSF IL-6, white blood cell count, and total protein starting 35 dpi with peak levels of these parameters coinciding with relapse parasitaemia. Brain immunohistochemical staining revealed an increase in brain glial fibrillary acidic protein expression indicative of reactive astrogliosis in infected animals which were euthanized in late-stage disease. The elevation of IL-6 in CSF which accompanied other HAT biomarkers indicates onset of parasite neuroinvasion and show potential for use as an adjunct late-stage disease biomarker in the Rhodesian sleeping sickness.

Relationship between the Prevalence of Ectoparasites and Associated Risk Factors in Free-Range Pigs in Kenya.

A cross-sectional study was undertaken to determine the prevalence of ectoparasites and possible risk factors in free-range pigs from 135 farms of Busia District, Kenya. Three hundred and six pigs were examined for presence of external parasites using standard parasitological methods. Data on management practices including housing and history of acaricide spraying were also collected. The ectoparasites found in the pigs were Haematopinus suis (96.1%), Sarcoptes scabiei (63.7%), and ticks (29.7%). The tick species included Rhipicephalus appendiculatus (70%), Boophilus decoloratus (31%), and Amblyomma variegatum (12%). The occurrence of the infestations was associated with age, being highest in sows (S. scabiei) and finishers (ticks and H. suis). Male pigs had highest prevalences of H. suis and ticks, while female pigs had highest prevalence of S. scabiei. The prevalence of the parasitic infestations was significantly (P < 0.05) associated with their origin being either lower (H. suis and S. scabiei) or higher (ticks) in pigs originating from divisions with high rainfall. Housed pigs had significantly (P < 0.05) lower prevalence of H. suis and ticks than those from households without pig housing. It is concluded that the free-range pigs have high prevalence of ectoparasites, and effective control strategies focussing on improved animal husbandry and acaricide use should be implemented.

Risk factors associated with occurrence of nematodes in free range pigs in Busia District, Kenya.

Nematode infections are a serious constraint to pig production, especially where free range pig keeping is practiced. This study investigated the epidemiology of nematodes in free range pigs in Busia District, Kenya. Three hundred and six pigs from 135 farms were sampled for faeces that were analysed for nematode eggs per gram (EPG) of faeces using the McMaster technique. The nematode eggs were also identified to genus and species based on morphology. A questionnaire on risk factors was also administered to the pig owners. The overall prevalence and mean nematode EPG were 84.2% and 2,355, respectively. The nematode eggs were identified as those belonging to Oesophagostomum spp. (75%), Strongyloides ransomi (37%), Ascaris suum (18%), Metastrongylus spp. (11%), Trichuris suis (7%) and Physocephalus sexalatus (3%). The prevalence of nematodes was positively correlated (p < 0.05) with the amount of rainfall in the division of the pigs' origin (all nematodes except S. ransomi). The prevalence of nematodes was also associated with the age of the pigs. A lower burden of nematodes was associated (p < 0.05) with a history of deworming (A. suum) and the provision of night housing (S. ransomi and Metastrongylus spp.). In conclusion, this study has provided information on nematode infections and the associated risk factors for free range pigs in Busia District, which can be used when implementing integrated control measures.

Proinflammatory cytokine expression in the early phase of Trypanosoma brucei rhodesiense infection in vervet monkeys (Cercopithecus aethiops).

A vervet monkey model of trypanosomiasis was used to study inflammatory cytokine responses in serum and cerebrospinal fluid (CSF). Gamma interferon levels were transiently up-regulated in serum between days 6 and 8 of infection, followed by a sustained up-regulation of tumor necrosis factor alpha (TNF-alpha) and soluble TNF receptor 1. At no time were these cytokines detectable in the CSF.