Modified Ziehl-Neelsen Staining Research Articles

Cryptosporidium and associated risk factors in local breeds of pigs kept in Donga Local Government Area Taraba State, Nigeria

Cryptosporidium, an enteric protozoan parasite, poses economic losses in animal production and has zoonotic potential. This study aimed to determine the prevalence of cryptosporidium and associated risk factors in local pig breeds within Donga Local Government Area of Taraba State, Nigeria. A cross-sectional study was conducted, and a total of 384 faecal samples were collected from pigs in eight sampling sites. The samples were examined using the modified Ziehl-Neelsen staining technique.The results showed an overall prevalence of 27.9% (107/384) for cryptosporidium oocysts. The prevalence was significantly higher in pre-weaned piglets (64%) compared to post-weaned (26.9%) and adult pigs (24.14%). Female pigs had a significantly higher prevalence (49.21%) than males (17.4%), and diarrhoeic pigs had a significantly higher prevalence (55.2%) than non-diarrhoeic (22.9%). The prevalence was higher in the rainy season (42.4%) than in the dry season (10.3%).The detection of Cryptosporidium spp. oocysts in the faeces of pigs suggests that these animals may serve as potential reservoirs and shedders of this protozoan parasite, contaminating the environment and serving as a source of infection to other animals and humans. Adequate personal and environmental hygiene, along with prompt veterinary care, should be intensified.

Identification of major protozoal enteropathogens causing calf diarrhea in dairy farms in and around Holeta Town, Oromia Special Zone, Ethiopia

Calf diarrhea can be caused by a variety of pathogens, including bacteria, viruses, protozoa, and intestinal parasites. Giardia, Eimeria, and Cryptosporidium are the most significant protozoan parasites and are all individually and  ollectively infectious. A case-series study was carried out in and around Holeta dairy farms, Oromia Special Zone, Ethiopia, from November 2017 to April 2018 to identify the main  protozoan enteropathogens from diarrheic calves. Samples were purposely collected from three dairy farms: Serkalem Dairy Farm (SDF),  Holeta Agricultural Research Centre (HARC), and Ada Berga Agricultural Research Centre (ABARC). A total of 93 fecal samples were taken from diarrheic calves of up to 4 months of age. Protozoan enteropathogens were identified using flotation and modified Ziehl Neelsen  staining methods. Cryptosporidium, Eimeria, and Giardia were detected in 44 (47.3%), 50 (53.8%), and 34 (36.6%) of the diarrheic fecal  samples examined, respectively. The findings indicated that there were 17 (18.27%) cases of Cryptosporidium, Eimeria, and Giardia as  mixed infections, 16 (17.2%) cases of Cryptosporidium and Eimeria, 5 (5.4%) cases of Cryptosporidium and Giardia, and 4 (4.3%) cases of  Eimeria and Giardia, compared to 13 (13.97%), 6 (6.45%), and 5 (5.4%) cases of Cryptosporidium, Eimeria and Giardia as single infections,  respectively. The selected farms had significant prevalence levels of three common protozoa: Eimeria, Giardia, and Cryptosporidium.  Further molecular research is required to identify the species and genotype levels of protozoal enteropathogens and related risk factors. 

Intestinal Parasitic Infections in People Referring to the Central Laboratory of Meshkin Shahr County, Ardabil Province, Iran.

Intestinal parasitic infections are still a considerable global public health problem. We aimed to determine the frequency of intestinal parasitic infections among people referring to the central laboratory of Meshkin Shahr City, Ardabil Province, Iran. In this cross-sectional survey, 460 fecal samples were collected randomly from persons referred to the central laboratory of Meshkin Shahr City, from January to June 2022. The samples were examined by direct wet-mount, Trichrome and modified Ziehl-Neelsen staining, formalin ethyl acetate sedimentation, and agar plate culture. The frequency of intestinal parasites was 15.7% (72 out of 460 cases), with some people with numerous intestinal parasites. The frequency of protozoan infections (13.9%) was higher than the helminthic infections (2.6%). Blastocystis spp. (8.1%) was the most prevalent detected intestinal protozoan. Entamoeba coli (5.7%), Dicrocoelium dendriticum (2.2%), Giardia lamblia (1.5%), Fasciola spp. (0.2%), and Hymenolepis nana (0.2%) were other detected parasites. In- spite of betterment of the health condition in Iran and reduction of parasitic infection, intestinal parasitic infections are still a considerable public health issue in some parts of Iran.

Pyroptosis Tuning in Intestinal Cryptosporidiosis via the Natural Histone Deacetylase Inhibitor Romidepsin.

Cryptosporidium is an opportunistic protozoan, with many species of cross-human infectivity. It causes life-threatening diarrhoea in children and CD4-defective patients. Despite its limited efficacy, nitazoxanide remains the primary anti-cryptosporidial drug. Cryptosporidium infects the intestinal brush border (intracellular-extracytoplasmic) and down-regulates pyroptosis to prevent expulsion. Romidepsin is a natural histone deacetylase inhibitor that triggers pyroptosis. Romidepsin's effect on cryptosporidiosis was assessed in immunocompromised mice via gasdermin-D (GSDM-D) immunohistochemical expression, IFN-γ, IL-1β and IL-18 blood levels by ELISA, and via parasite scanning by modified Ziehl-Neelsen staining and scanning electron microscopy (SEM). Oocyst deformity and local cytokines were also assessed in ex vivo ileal explants. Following intraperitoneal injection of romidepsin, oocyst shedding significantly reduced at the 9th, 12th and 15th d.p.i. compared with infected-control and drug-control (nitazoxanide-treated) mice. H&E staining of intestinal sections from romidepsin-treated mice showed significantly low intestinal scoring with marked reduction in epithelial hyperplasia, villous blunting and cellular infiltrate. SEM revealed marked oocyst blebbing and paucity (in vivo and ex vivo) after romidepsin compared with nitazoxanide. Regarding pyroptosis, romidepsin triggered significantly higher intestinal GSDM-D expression in vivo, and higher serum/culture IFN-γ, IL-1β and IL-18 levels in romidepsin-treated mice than in the control groups. Collectively, in cryptosporidiosis, romidepsin succeeded in enhancing pyroptosis in the oocysts and infected epithelium, reducing infection and shifting the brush border towards normalisation.

Prevalence and risk estimates of Cryptosporidium oocysts infection associated with consumption of raw-eaten vegetables in Maiduguri metropolis LGAs, Northeast Nigeria

Cryptosporidium is one of the most important protozoan parasitic pathogens, and it is a common cause of diarrhoea in humans, domestic animals, and wild vertebrates and has serious public health threats. A cross-sectional study was designed to determine the prevalence of Cryptosporidium oocysts in raw-eaten vegetables in Maiduguri Metropolitan Council (MMC) and Jere Local Government Areas (LGAs). A total of 400 samples were collected from four (4) different locations, namely Tashan Bama, Gomboru, and Monday Markets (n = 100), while fifty (n = 50) each from 202-Vegetable-Vendors and Unimaid Commercials. A total of 16 visits were conducted in all the sampling areas (twenty-five samples per visit). The Cryptosporidium oocysts were detected using the Modified Ziehl–Neelsen Staining Technique. The locations, sources, and types of raw-eaten vegetables were also assessed. The oocysts were confirmed (100×) as bright pink spherules. Data generated were analyzed using IBM-SPSS V23.0, and p ≤ 0.05 was considered significant. Out of the total samples (n = 400) analyzed, cabbage appeared to have the highest number of 10 (12.5%) of Cryptosporidium oocysts detected, while Tomato and garden egg had 9 (11.3%) and 1 (1.2%), respectively. There was a statistically significant association (χ2 = 12.5, P = 0.014) between the presence of Cryptosporidium oocysts in raw-eaten vegetables and vegetable types. Among the sources of the vegetables sampled, Alau had the highest number of Cryptosporidium oocysts, 15 (12.5%), followed by Kilari-Abdullahi and Zabarmari sources with 4 (10.0%) and 4 (5.0%), respectively. However, Jetete appeared to have the least number 2 (2.5%) of oocysts, and there was a statistically significant association (χ2= 10.4, P = 0.034) between the presence of Cryptosporidium oocysts and the sources of vegetables and fruits. The study concludes that the raw-eaten vegetables sampled from Maiduguri Metropolis were contaminated with Cryptosporidium oocysts. The study recommends that all raw-eaten vegetables should be from cleaned sources and washed before consumption. Consumers should be enlightened on the hygienic measures in the food chain in line with the Hazard Analysis and Critical Control Points (HACCP) principles.

Enteric Protozoan Parasitosis and Associated Factors among Patients with and without Diabetes Mellitus in a Teaching Hospital in Ghana.

Enteric protozoa infections (EPIs) could worsen clinical outcomes in patients with diabetes mellitus and therefore requires prompt and accurate diagnosis and attention. This study aimed to determine the burden of EPIs and their associated factors among patients with and without diabetics at the Komfo Anokye Teaching Hospital (KATH) in Ghana. Again, the diagnostic performance of parasitological techniques routinely used for diagnosis was assessed. A total of 240 participants (made up of 140 patients with diabetes and 100 patients without diabetes) were recruited into the study by simple random sampling from November 2020 to May 2021. Stool samples of participants were collected, along with their demographic information, and examined using the saline direct wet mount (DWM), formol-ether concentration (FEC), and modified Ziehl-Neelsen staining (ZNS) techniques for the presence of enteric protozoans. Enteric protozoa were found among 62/140 (44.3%) diabetic patients and 13/100 (13.0%) nondiabetic patients. The predominant protozoa identified were Cryptosporidium spp. (17.86%) among patients with diabetes and Blastocystis hominis (7.0%) among patients without diabetes. EPI was associated with diabetes mellitus status (AOR = 3.48, 95% CI, 1.55-7.79), having diabetes for more than five years (AOR = 3.83, 95% CI, 1.65-8.86) and having comorbidity (AOR = 2.93, 95% CI, 1.33-6.45). The FEC technique had the highest sensitivity (100.0%), specificity 94.3% (95% CI, 91.35-97.22), and accuracy 95.0% (95% CI, 88.54-98.13) when compared to other techniques for diagnosis. EPIs are a significant health problem among patients with diabetes at KATH, and therefore antiparasitic drugs should be included in their treatment protocols for better health outcomes. Again, the FEC technique has demonstrated better performance in detecting EPIs and is therefore recommended to achieve early and accurate diagnosis of EPIs.

Detection and molecular characterization of Cryptosporidium spp. in dairy calves in Lisbon and Tagus Valley, Portugal

Cryptosporidium is a protozoan parasite with worldwide distribution, infecting a wide range of hosts with some zoonotic species. Calves have been identified as one of the most common reservoirs of this parasite. However, little is known about the genetics of Cryptosporidium in calves in Portugal. This study aimed to molecularly characterize infections of Cryptosporidium in pre-weaned calves from the Lisbon and Tagus Valley (LTV) in Portugal. Fifty-two samples were collected from calves from eight dairy and two beef farms in LTV, Portugal. Cryptosporidium oocysts were detected by Modified Ziehl-Neelsen staining (MZN) and direct immunofluorescent assay (DFA). MZN and DFA revealed the presence of Cryptosporidium oocysts in 40.4% (21/52) and 67.3% (35/52) samples, respectively. Positive samples were analyzed by PCR-RFLP of the 18 s rRNA gene for species identification. DNA amplification of the 18S rRNA gene was successful for 88.6% (31/35) of samples. Cryptosporidium parvum was identified in 96.8% (30/31) of the samples, and from one sample Cryptosporidium bovis was identified. Cryptosporidium parvum positive samples were subtyped by sequencing the PCR product of a partial fragment of the 60 kDa glycoprotein (gp60) gene. Subtype analysis of the C. parvum isolates revealed that all isolates belonged to subtype family IIa. Four subtypes were recognized within this subtype family, including the hyper-transmissible IIaA15G2R1 subtype that is the most frequently reported worldwide (27/30), IIaA14G2R1 (1/30), IIaA16G2R1 (1/30) and IIaA19G2R1 (1/30). To our knowledge, this is the first report of C. bovis, and C. parvum subtypes IIaA14G2R1 and IIaA19G2R1 in cattle in LTV, Portugal. The presence of the zoonotic C. parvum subtype in this study suggests that pre-weaned calves are likely to be a significant reservoir of zoonotic C. parvum, highlighting the importance of animal-to-human infection transmission risk. Further molecular studies are required to better understand the epidemiology of cryptosporidiosis in Portugal.

Molecular Characterization of Cryptosporidium Oocysts in under-Five Children with Diarrhoea in Ile-Ife, Nigeria

Aim: Cryptosporidiosis is the fifth leading cause of diarrhoea-related death globally among under-five children. This study was conducted to identify and characterize oocysts of Cryptosporidium in under-five children with diarrhoea in Ile-Ife.
 Study Design: It was a case-control study.
 Place and Duration of the Study: Department of Medical Microbiology and Parasitology, Obafemi Awolowo University between April, and October 2019.
 Methodology: Ethical approval was obtained for this study. Consent was also obtained from the parents of the children. Stool samples from 53 children with diarrhoea (cases) and 94 children without diarrhoea (controls) under five years of age were collected. A structured proforma was used to obtain relevant information from the parents of the children. Stool samples were collected and examined macroscopically for consistency, appearance, and colour. Modified Ziehl-Neelsen staining was carried out on all the stool samples. The samples positive for Cryptosporidium oocysts by staining were genotyped by nested polymerase chain reaction and confirmed by sequencing of their 18S rRNA genes. Data generated were analyzed using descriptive and inferential statistics with SPSS software version 20 and STATA 15. 
 Results: The prevalence of cryptosporidiosis among the study participants was 23.1% with rates of 43.4% (23) and 11.7% (11) for diarrhoeic and non-diarrhoeic children respectively. Of the 34 stool samples analysed, 6 (17.6%) were amplified by nested PCR and 4 (11.8%) were identified by sequencing. The species of Cryptosporidium identified were Cryptosporidium parvum (75%) and Cryptosporidium bovis (25%).
 Conclusion: In conclusion, Cryptosporidiosis is prevalent among under-five children with Cryptosporidium parvum and Cryptosporidium bovis as the infecting species.

[Ocorrência de Cryptosporidium spp. em bezerros de rebanhos leiteiros do norte de Antioquia, Colômbia]

ABSTRACT Cryptosporidium spp. are important enteropathogen protozoan parasites that infect humans and other animals throughout the world. Cryptosporidiosis in cattle industry leads to considerable economic losses due to diarrhea, dehydration, growth retardation, weight loss, and possibly death, however, data on the occurrence ofCryptosporidiumspp. in cattle in Colombia are limited. Therefore, this study aimed to investigate the occurrence and possible factors associated to the excretion of Cryptosporidium spp. oocyst in pre-weaned calves from dairy farms in Northern Antioquia, Colombia. In addition, Sheather’s sugar floatation (SSF), and Modified Ziehl-Neelsen staining (MZN) methods were compared. A total of 41 fecal samples were collected from calves between 1 and 92 days of age of which 23 were positive (56.1%). Crossbreed calves were nine times less susceptible than purebred dairy cattle to excretion of Cryptosporidium spp. oocysts (OR=0.10). MZN was the best technique for the detection of oocysts in fecal samples, however, the mean number of days to detect cryptosporidial oocysts was lower for the SSF method. Cryptosporidium is widespread among calves under 2 months of age in dairy herds of Northern Antioquia, although further investigations considering a greater number of farms and animals are necessary.

Dynamics of protozoal excretion in the faeces of calves during the first 28 days after arrival at the fattening farm indicate infection before regrouping and show poor temporal correlation with diarrhoea

BackgroundCalves in dairy cattle production in Switzerland are transported to a fattening farm at the age of 3–5 weeks, and frequently suffer from diarrhoea within the first 14 days after arrival. To characterise the role of intestinal protozoa in this, we investigated the excretion dynamics of Eimeria, Cryptosporidium and Giardia during the first 28 days after the arrival and regrouping of calves at fattening farms.MethodsA total of 610 faecal samples from 122 calves (mean age 37.3 days; mean body weight 79.8 kg) were collected on seven different fattening farms during the first 28 days after the arrival and regrouping of the animals. The farms were visited between January and April (cold season; n = 4) and between June and August (warm season; n = 3). The samples were collected rectally on days 1, 4, 7, 14 and 28, assessed for consistency, and analysed using the McMaster method for quantitative determination of the number of Eimeria oocysts per gram of faeces (OPG), flotation for morphological differentiation of the unsporulated Eimeria oocysts, a concentration method for the semi-quantitative determination of Giardia cysts, and modified Ziehl–Neelsen staining for semi-quantitative determination of Cryptosporidium oocysts.ResultsOverall, 50.8% (62/122) of the animals had diarrhoea during the study period. However, the faecal excretion of protozoal pathogens was neither associated with diarrhoea nor with body weight gain of the animals. Altogether, 90.2% (110/122) of the calves were Eimeria positive. Eimeria zuernii was excreted by 51 (41.8%) and Eimeria bovis by 68 (55.7%) animals. In the warm season more animals tested positive for Eimeria and OPGs were higher than in the cold season. There was no correlation between the age of the calves and the OPG values. Overall, 64.8% (79/122) of the calves excreted Eimeria oocysts within the first 7 days, indicating that they had been infected with the parasite on the dairy farm of origin. Eighty-nine calves (73.0%) excreted Giardia cysts, with more positive animals in the cold (80.3%) compared with the warm season (64.3%). Only Giardia duodenalis assemblage E was identified. Cryptosporidium oocysts were microscopically detected in 14 animals (11.5%) on five farms. Cryptosporidium spp. were present in a total of 12 animals, i.e. Cryptosporidium parvum in nine, Cryptosporidium ryanae in two, and Cryptosporidium bovis in one animal.ConclusionsA better understanding of the temporal dynamics of protozoal infections in calves is helpful for the implementation of appropriate measures to protect the health of these animals at a critical phase in their lives. Our results indicate that factors other than those examined in the present study contributed to the onset of diarrhoea in the calves.Graphical abstract

Diagnostic Performance of Toluidine Blue Stain for Direct Wet Mount Detection of Cryptosporidium Oocysts: Qualitative and Quantitative Comparison to the Modified Ziehl-Neelsen Stain.

(1) Background: The wet mount staining technique is a simple, economical, and rapid method for detecting parasitic stages. The objective of the current study was to evaluate wet mount diagnostic stains for Cryptosporidium infection in human faecal samples and to compare the sensitivity and qualitative performance of toluidine blue (TolB) and modified Ziehl-Neelsen (mZN) stain. (2) Methods: The collection, purification, and molecular amplification of Cryptosporidium oocysts were performed. TolB, malachite green, trypan blue, and crystal violet were evaluated qualitatively to diagnose Cryptosporidium oocysts. The outperforming stain was compared to mZN using a quantitative evaluation and qualitative scoring system. (3) Results: The oocysts of Cryptosporidium parvum were effectively purified and utilized for spiking. TolB was the most effective diagnostic stain among wet mount stains for detecting Cryptosporidium oocysts. TolB outperformed mZN in terms of sensitivity, with 100% versus 79% at a concentration of 104 and 49% versus 23% at a concentration of 102. TolB had the most favourable qualitative stain characteristics against mZN; however, sample freshness and being a temporary stain were crucial considerations. (4) Conclusions: This study emphasizes that TolB, as a routine wet mount technique for screening Cryptosporidium infection, will provide a more sensitive and faster method than mZN staining.

Spectrum of Biopsy Proven Skin Lesions in Armed Forces Population: A Tertiary Care Hospital Experience

Abstract Background: Skin disorders are common as skin is constantly exposed to various chemicals, ultraviolet rays, climate, etc., making it prone to damage leading to various diseases. The spectrum of skin disorder lesions varies according to geographical distribution, gender, and age. A large number of skin lesions can be diagnosed clinically, but histopathological examination is necessary for accurate diagnosis and proper management. Literature shows few such studies in India, but no such study could be found for the Indian Armed Forces population. The aim of our study was to understand the pattern of skin diseases in Armed Forces population. Materials and Methods: All skin biopsies received between January 2022 and December 2022 were included in the study. Two hundred skin biopsies received were examined for various skin ailments. The skin biopsies were processed, sectioned, and stained with hematoxylin and eosin and wherever required special stains like modified Ziehl–Neelsen stain. The histopathological data were retrieved and reviewed. The data were collected and entered into SPSS 20.0 for statistical analysis. Results: Out of the two hundred cases studied, the most common group of disorders were of noninfectious nonneoplastic category (55%), followed by benign neoplasms (25%) and infectious category (18%), with least cases belonging to malignant category (2%). Commonly involved site in the body was upper limbs. The most common disorder found in males was psoriasis and in females was lobular capillary hemangioma. Conclusion: Understanding spectrum of histopathological spectrum of skin disorders will aid in identifying and managing different disorders, effectively providing high-quality care to the patient.

PARASITIC CONTAMINATION OF VEGETABLES IN SELECTED LOCAL MARKETS IN ADEN GOVERNORATE, YEMEN

Fresh and raw vegetables may play a significant role in transmission of intestinal parasitic infections to humans. Therefore, this study was conducted to estimate the level of parasitic contamination in vegetables which are consumed raw in Aden governorate, Yemen. This is a descriptive analytic cross-sectional study in which six local markets were randomly selected. A total of 216 fresh vegetable samples including parsley (Petroselinum sativum), watercress (Nasturtium officinale), lettuce (Lactuca sativa), leek (Allium porrum), green onion (Allium cepa), and tomatoes (Solanum Lycopersicum) were purchased from the selected markets and analysed using direct and iodine wet mount smear preparation. Modified Ziehl–Neelsen staining and Zinc sulphate flotation method were also used. Smears were examined by under the light microscope. Out of 216 vegetable samples, 166 (76.9%) were contaminated with one or more parasites. Parsley was the most contaminated (97.2%), followed by watercress (94.4%), lettuce (88.9%), leek (80.6%), green onion (77.8%), whereas the least contamination rate was in tomato (22.2%). Statistically significant difference in parasitic contamination rate was found between different types of vegetables (P < 0.05). The most common parasite was Ascaris lumbricoides 75 (24.4%), followed by cyst of Entamoeba histolytica/dispar 64 (20.9%), and Entamoeba coli 52 (16.8%). Significant difference was reported in prevalence rate between different parasitic species (????< 0.05). Vegetable samples collected from Dar Saad market, show the highest contamination rate (94.4%) while vegetables collected from Al Mansura market showed the lowest contamination rate(50.0%). However, the parasitic contamination rates were not significantly different for samples collected from different markets. This study highlighted the importance of raw vegetables as the potential source of transmission for intestinal parasites to humans. Prevention methods such as proper washing or cooking of vegetables before consumption should be conveyed to consumers. In addition, comprehensive health education and hygienic practices, including washing hands after handling vegetables, should be provided to sellers and consumers.

Risk Factors of Cryptosporidium spp. Infection Through Drinking Water Sources in Farming Communities in Sukowono District Jember Regency

Cryptosporidiosis is a zoonotic, emerging water-borne and protozoan disease. This parasite infects the digestive and respiratory tracts of human and various animals. This study aims to determine the risk factors of Cryptosporidium spp. infection through drinking water sources in farming communities in Sukowono District, Jember Regency and using an observational analytic study with a cross-sectional approach. The research sample was reviewed using observational sheets and stool samples with concentration method and using modified Ziehl-Neelsen staining on research subjects’ samples based on inclusion and exclusion criteria totaling 52 people. The results of this study indicate the characteristics of the research subjects in Sukowono District, Jember Regency, dominated by female gender (57.7%), adult age category (42.3%), elementary school graduate (53.8%), and identified as a farmer or rancher (34.6%). The examination of stool samples showed Cryptosporidium spp. an infestation in 1 (one) person. The results of the bivariate analysis using the Fisher's Exact alternative test obtained p of 0.601 (p>0.05). The conclusion is that the majority of research subjects use unhealthy well water as drinking water source, there is one person infected, also all factors from drinking water sources are not at risk of this parasitic infection in Sukowono District, Jember Regency.

The Intensity of Infection and Public Health Perception of Potentially Zoonotic Intestinal Parasites of Dogs in Kwara Central, Nigeria

Background: The close relationship between dogs and humans and the possibility of intestinal parasite transmission from dogs to humans calls for frequent assessment of these potential zoonotic intestinal parasites in dogs and the possibility of their transmission to humans. Objectives: This study aimed to determine the presence, intensity of infection, and public health perception of potentially zoonotic intestinal parasites in dogs of Kwara Central, Nigeria. Methods: The study was conducted in 28 locations in Kwara Central Senatorial District of Kwara State, Nigeria. Three hundred and five healthy dogs were sampled. Two hundred and thirty respondents (dog owners or handlers) were questioned using a well-structured questionnaire containing open-ended and closed-ended questions. Fecal samples from the sampled dogs were subjected to the direct fecal smear technique, simple fecal centrifugation flotation technique, formalin-ethyl acetate concentration technique, and the modified Ziehl-Neelsen staining technique. Oocysts or eggs per gram of feces were counted using the modified McMaster technique. Results: Seven different intestinal parasites (Ancylostoma spp., Cryptosporidium spp., Dipylidium caninum, Isospora spp., Strongyloides stercoralis, Toxocara spp., and Uncinaria stenocephala) were detected, with a prevalence ranging from 2.30% to 25.25%. Of these parasites, 6 were zoonotic. The mean intensities of infection were 91.43 eggs per gram (EPG) for D. caninum¸ 96.52 EPG for S. stercoralis, 129.36 ±28.12 oocysts per gram (OPG) for Isospora spp., 165.17±19.88 for Toxocara spp., 240.00±44.42 for EPG for U. stenocephala, and 303.64±31.83 EPG for Ancylostoma spp. Some dog owners and handlers were not cautious about possible zoonotic parasite transmission from dogs. Conclusion: Zoonotic intestinal parasites of dogs are present and prevalent in Kwara Central, Nigeria. There is a need to educate the public on the possibility of zoonotic parasite transmission to humans.

Assessment of Co-Infection in Tuberculosis Patients Prior to their Enrollment to the Direct Observed Treatment (Dot) Program in Buea, Limbe, and Douala Tuberculosis Treatment Centers

Introduction: Co-infections during Tuberculosis infection are among the leading causes of morbidity and mortality worldwide. These infections are a major public health problem among patients in most developing countries, including Cameroon. One of the important public problems in coendemic areas is co-infection with TB in humans. Moreover, there is no adequate research hitherto that provides information on the level of co-infection in Tuberculosis patients. Therefore, this study aimed to assess the prevalence and intensity of co-infections in newly diagnosed Tuberculosis patients prior to their enrollment to the Direct Observed Treatment (DOT) program in Buea, Limbe and Douala Tuberculosis treatment centers. Methods: This cross-sectional study was conducted from March to November 2020 using a purposive sampling technique. A fresh sputum sample was collected, processed, and examined through a smear microscopy and culturing technique. Blood samples were collected through finger pricking for the detection of blood parasites using thin and thick blood smears. Stool samples were also collected and processed using a direct wet mount, and Kato-Katz and Modified Ziehl Neelsen staining methods. The data was analyzed using SPSS version 20.0. A Chi-square test was done to assess the prevalence of co-infection among all TB patients recruited for this study. Results: A total of 200 newly diagnosed TB-positive patients were recruited from the three study sites. Out of the total participants recruited, 29.0% were diagnosed positive for blood parasites, 2.0% for intestinal parasites and 15.0% for viral infections, bringing the overall prevalence of co-infection to 46.0%. When specific infections were considered, malaria, filaria, intestinal parasites, HIV and HBV constituted 28.0%, 1.0%, 2.0%, 14.5% and 0.5% respectively. When dual-infection was considered, the results were 0.5% malaria/filaria, 1.5% malaria/intestinal parasites and 2.0% malaria/HIV. The mean parasite densities for all samples diagnosed positive for malaria, filarial and intestinal parasites were 107.0 ± 338.8 parasites/uL, 25.7 ± 356.4 mf/mL and 85.0 ± 85.4 EPG, respectively. Conclusion: The results indicate that there is an overlap of co-infections in TB patients with their interactions expected to lead to complications and difficulty when managing the TB patients, calling for more research and recommending more assessment for co-infection in TB patients before starting treatment.

Occurrence of Gastrointestinal Parasites among Patients Attending Designated Tuberculosis Clinics in Rivers State, Nigeria

Diseases of gastrointestinal parasites remain a major public health concern. This study was aimed at determining the occurrence of gastrointestinal parasites among patients attending designated tuberculosis clinics in Rivers State, Nigeria. The clinics included Chest clinic, Rumuigbo, General hospital, Ahoada and Meridian hospital. The study was conducted between July 2020 and August 2021. Stool and blood samples were collected from 1288 participants and examined for the presence of ova, cysts, oocyst and trophozoites of gastrointestinal parasites using formol-ether concentration and Modified Ziehl-Neelsen staining techniques. ELISA was used to examine the blood samples for the presence of Cryptosporidium spp. and Giardia lamblia. Out of the 1288 patients examined, 580 (45%) were infected and this was statistically significant (p<0.05). Gastrointestinal parasites identified included Ancyclostoma spp., Ascaris lumbricoides, Cryptosporidium spp, Entamoeba histolytica, and Giardia lamblia. Cryptosporidium spp. was the most prevalent (p<0.05). Females had a higher prevalence (55.9%) than males (44.1%), and the age group 11-20 years were most infected. The occurrence of gastrointestinal parasite infection was higher in the wet season (p<0.05). Sources of drinking water and housing types were risk factors that had a significant influence on the infection (p<0.05). The gastrointestinal parasite was not influenced by the occupation, as well as the educational levels of the participants. The study showed that tuberculosis patients were more infected than non-tuberculosis patients, and occurrence was higher in rural areas than in urban areas; both were significant (p<0.05). Detection of Giardia lamblia and Cryptosporidium using non-ELISA and ELISA techniques were not statistically significant (p>0.05). The study showed a higher prevalence of infection; therefore, there is a need to enhance health education on the mode of transmission of gastrointestinal parasites and improve on personal and environmental hygiene of tuberculosis patients and the population as a whole; to reduce the occurrence of infection

The Presence of Potentially Pathogenic Protozoa in Lettuce (Lactuca sativa) Sold in Markets in the Central Peruvian Andes.

Peru is currently one of the world’s leading culinary destinations, whose world-renowned cuisine uses vegetables, mainly lettuce, as frequent ingredients. Vegetable consumption is promoted worldwide as a part of a healthy diet. However, vegetables, more frequently lettuce, have been implicated as a vehicle of infection for several foodborne parasites. This study aimed to determine the presence of potentially pathogenic parasites in lettuce marketed in the Central Andes of Peru. A total of 75 lettuce samples were collected from the two largest wholesale markets and the main open-air market in Jauja province, in the central Peruvian Andes. The province of provenance (coast vs. highlands), lettuce variety (“curly-leaf”, “iceberg”, and “butter”), and type of market were recorded. The samples were microscopically examined for detection of parasitic life forms using standard parasitological methods including direct slide smear, Lugol’s iodine staining, and Modified Ziehl−Neelsen staining. The overall positivity of parasitic contamination in lettuces was 45.3% (CI 95%: 34−56.6%). Cryptosporidium spp., Isospora belli, Giardia lamblia, Balantidium coli, and Entamoeba spp. were detected in twenty-six (34.7%), six (8%), four (5.3%), two (2.7%), and two (2.7%) lettuces, respectively. I. belli was found in a significantly (p < 0.01) lower proportion in the “butter” variety, and significantly (p < 0.05) higher contamination with G. lamblia was found in lettuce sold at the open-air market. B. coli, G. lamblia, and E. histolytica/E. dispar/E. moshkovskii were detected only in lettuce from the highlands (Tarma province). This study provides important data for health authorities to develop food safety programs. This information is also of interest to the international community because of the increased visibility that Peru has gained as a tourist destination.

Epidemiological studies of gastrointestinal parasites infecting dogs in Kwara Central, North Central, Nigeria

Dogs are the most cosmopolitan pets of humans and as such a means of transmitting zoonotic parasites to their owners. This study was designed to investigate the diversity, prevalence, pattern of infection, intensity of infections, and the risk factors associated with gastrointestinal parasites of dogs in Kwara Central, North Central, Nigeria. Three hundred and five clinically healthy dogs were sampled. Faecal samples were subjected to the direct smear, simple faecal centrifugation flotation, formol-ether concentration, and the Modified Ziehl-Neelsen staining techniques. Oocysts/eggs per gram of faeces were counted using the modified McMaster technique. Data were analysed using univariate logistic regression, multivariate logistic regression, and the one-way analysis of variance (ANOVA). A p -value of < 0.05 was considered significant for all analyses. One hundred and sixty-six dogs were positive for at least one species of gastrointestinal parasite, representing 54.43% (95% CI: 44.81 – 59.96) of the sampled population. The study identified Cystoisospora species (15.41%), Cryptosporidium species (25.25%), Ancylostoma species (25.25%), Toxocara canis (19.02%), Strongyloides stercoralis (7.54%), Uncinaria stenocephala (6.89%), and Dipylidium caninum (2.30%) as the gastrointestinal parasites infecting dogs in the study area. Coinfection with more than one species of gastrointestinal parasites was a common finding in dogs. The intensity of Cystoisospora spp. among infected dogs ranged between 40 and 980 oocysts per gram of faeces, while that of helminth parasites was 40 – 1560 eggs per gram of faeces. Age, sex, breeds, body condition score, presence of ticks on dogs, the purpose of keeping dog(s), types of housing, types of feed consumed, vaccination status, and treatment with antiparasitics were predators associated with the prevalence and intensity of gastrointestinal parasites infections. Due to the zoonotic nature of most of the encountered gastrointestinal parasites, there is need for regular antiparasitic treatment, proper dog management, and adequate personal hygiene to prevent zoonosis.

Endoparasites of Leopardus pardalis (Carnivora, Felidae) rescued in the State of Rio de Janeiro.

Wild cats play an important role as top predators in the food chain and act as ecosystem regulators. However, in recent decades, many studies have demonstrated the potential effects of parasitic diseases on wild carnivore populations, including cats. This study reports on the endoparasites found in an injured and rescued specimen of Leopardus pardalis in the state of Rio de Janeiro. Fecal samples were collected and processed using five coproparasitological techniques: a simple flotation centrifugation, a zinc sulfate flotation centrifugation, a formalin-ether sedimentation centrifugation, a conical centrifuge tube technique, and a modified Ziehl-Neelsen staining technique for fecal smears. Helminth eggs belonging to the families Trichuridae and Diphyllobothriidae and the genus Toxocara were found in both sedimentation flotation techniques. Protozoan oocysts belonging to the genus Cryptosporidium were identified by modified Ziehl-Neelsen staining. These findings show that ocelots can harbor potentially zoonotic and pathogenic endoparasites. Further studies on the helminths and protofauna of these animals are necessary.