Coccidial Infection Research Articles

Probiotic Enterococcus faecalis surface-delivering key domain of EtMIC3 proteins: immunoprotective efficacies against Eimeria tenella infection in chickens.

Enterococcus faecalis MDXEF-1 strain was used as host bacteria to deliver surface-displayed key domains BC1 and C4D of the Eimeria tenella microneme 3 (EtMIC3) protein, BC1 fusion to dendritic cell (DC) targeting peptides (DCpep) (DC-BC1) and DC-C4D, respectively. The expression of target proteins in MDXEF-1/BC1-CWA, MDXEF-1/DC-BC1-CWA, MDXEF-1/C4D-CWA, and MDXEF-1/DC-C4D-CWA was verified. The systemic immune responses induced by oral immunization with recombinant E. faecalis were recorded. The immunized chickens were challenged with 4.0 × 104 E. tenella-sporulated oocysts. The immune protections were evaluated by calculating average weight gain and oocyst reduction rate, observing gross and histopathological changes in the cecal tissues, and quantifying levels of inflammatory cytokines in the cecal tissues. The results showed that proteins BC1, C4D, DC-BC1, and DC-C4D were expressed on the surface of E. faecalis, respectively. Oral immunizations with recombinant E. faecalis, especially MDXEF-1/DC-BC1-CWA and MDXEF-1/DC-C4D-CWA, stimulated higher levels of antigen-specific humoral and intestinal mucosal immune responses, higher levels of Th1, Th2, and Th17-type cytokines, and higher proliferation of peripheral blood lymphocytes than phosphate-buffered saline (PBS) and vector control groups (P < 0.01). The groups immunized with four recombinant E. faecalis showed better protective effects against homologous infection than the vector control and infection control groups, and the MDXEF-1/DC-BC1-CWA group displayed the best effects. These results demonstrated that probiotic E. faecalis delivering DCpep fused with the key domain of the EtMIC3 protein could be a potential approach for the prevention of Eimeria infection. IMPORTANCE Avian coccidiosis caused by Eimeria brings huge economic losses to the poultry industry. Although live vaccines and anti-coccidial drugs were used for a long time, Eimeria infection in chicken farms all over the world commonly occurred. The exploration of novel, effective vaccines has become a research hotspot. Eimeria parasites have complex life cycles, and effective antigens are particularly critical to developing anti-coccidial vaccines. Microneme proteins (MICs), secreted from microneme organelles located at the parasite apex, are considered immunodominant antigens. Eimeria tenella microneme 3 (EtMIC3) contains four conserved repeats (MARc1, MARc2, MARc3, and MARc4) and three divergent repeats (MARa, MARb, and MARd), which play a vital role during the Eimeria invasion. Enterococcus faecalis is a native probiotic in animal intestines and can regulate intestinal flora. In this study, BC1 and C4D domains of EtMIC3, BC1 or C4D fusing to dendritic cells targeting peptides, were surface-displyed by E. faecalis, respectively. Oral immunizations were performed to investigate immune protective effects against Eimeria infection.

Malaria, HIV, and Intestinal Coccidian Parasites Mixed-Infections in Adult Patients in the Fundong Health District, Northwest Region, Cameroon

Background: Malaria and intestinal coccidian parasites are both protozoan and their interaction in co-infected patients is still not clear. Patients infected with both malaria and coccidian parasites experience diarrhoea, gastro-intestinal and health problems that maybe life threating. We studied opportunistic intestinal coccidian parasites (OIPs) and the human immunodeficiency virus (HIV) in adult patients with malaria in Fundong Health District, Northwest of Cameroon. The objectives was to determine prevalence, and distribution of the diseases in adult population. &#x0D; Methods: A prospective cross-sectional study carried out between April and December 2022. Malaria patients were identified by the presence of the Plasmodium parasite in Giemsa blood-stained films. HIV status was determined using the rapid diagnostic test (RDT). Stool samples were subjected to wet preparation and formol-ether concentration technique to detect intestinal coccidian parasites. Pearson’s Chi-Square (χ2) and binary logistic regression were performed as part of the statistical analysis. Statistical significance was set a P-value&lt;0.05.&#x0D; Results: Three hundred and thirty (330) adult patients all infected with malaria took part in the study, 115(35%) males and 215 females (65%). The adult were of age 21 years and above. The mean age of participant was 37.02(±15.235) years. Malaria co-infection with coccidian parasites was observed in 19.4%. Malaria co-infection with other gastro-intestinal intestinal parasite capable causing pathological conditions in patients was 5.5%. The study showed the prevalence of malaria co-infections with HIV was 8.2%. The mixed infections observed were domestically acquired (57.8%) as well as travel related (46.2%). Fever was most reported in severe malaria 315/330 (95%), followed by those who reported abdominal pains 85/330 (26%), while diarrhoea was reported in 35/330 (11%) of the study participants. Fever, and abdominal pain significantly associated with malaria status (simple or severe malaria) p-value=, P-value= respectively (5.915) 0.015, P-value= (8.3) 0.004, as well as the HIV status P-value= (11.798) 0.003. The odds of developing severe malaria from fever was 3.533(CI: 1.204-10.366) times higher compared to those who did not have fever. The odds of developing severe malaria from abdominal pain was 0.420(CI: 0.230-0.767) less. Results showed living with child&lt;2 years, water treatment methods, material use in hand washing, predicted the probability of coccidian infection in the final logistic regression model accounting for 23.6% of the variance in the coccidian infections was explained by the model.&#x0D; Conclusion: Prevalence of malaria, and opportunistic intestinal coccidian infections in adult patients was significantly influenced by migration, seasonal variation and the individual’s immune status. Routine clinical practice have often ignore the investigation of possible mixed infections especially in persons with compromised immunity including HIV. Screening patients suspected of malaria should include investigation for opportunistic intestinal coccidian parasites especially for HIV and other vulnerable populations. An integrated prevention and control strategy need to be considered.

Graded levels of Eimeria infection linearly reduced the growth performance, altered the intestinal health, and delayed the onset of egg production of Hy-Line W-36 laying hens when infected at the prelay stage

The aim of this experiment was to investigate how different levels of Eimeria infection affect the performance, intestinal health, oxidative status, and egg production of Hy-Line W-36 pullets and laying hens. Three hundred and sixty Hy-Line W-36 pullets, aged 15 wk, were randomly distributed into 5 treatment groups, each comprising 6 replicates and a nonchallenged control. At 15 wk, pullets were inoculated with different levels of mixed Eimeria species as high-dose, medium-high, medium-low, and low-dose treatments. The growth performance and average daily feed intake (ADFI) were measured from 0- to 18-days postinoculation (DPI), whereas hen day egg production (HDEP) was recorded from wk 19. The markers of gastrointestinal health and oxidative status were measured at 6 DPI, 14 DPI, and 23 wk of age. The findings revealed a significant linear reduction in growth performance in response to increasedEimeriachallenge dosage on 6 and 14 DPI (P < 0.0001,P-L < 0.0001). An interaction between thegraded level of Eimeriainfection and DPI was observed for ADFI. The challenged pullets showed a reduction in ADFI starting at 4 DPI, which persisted until 14 DPI, when ADFI recovered back to normal. The most significant drop in feed intake was observed in 6 DPI in all the Eimeria-infected groups. The markers of gastrointestinal health (gastrointestinal permeability and tight junction proteins) were upregulated in challenged pullets because of infection, whereas the relative mRNA expression of key nutrient transporters was downregulated following infection on 6 and 14 DPI (P < 0.05). As a result of an infection on 6 DPI, the oxidative equilibrium was shifted toward the oxidative stress, and at the same time, upregulation of proinflammatory and inflammatory cytokines was observed (P < 0.05). An interaction between the Eimeria challenge dosage and bird age was observed for HDEP (P=0.0427). The pullets infected with Eimeria started to lay eggs later than the Control birds. However, the HDEP of the challenged groups became similar to Control only at wk 22, 3 wk after laying eggs. In conclusion, coccidiosis reduced growth performance, altered gastrointestinal health, induced oxidative stress, and delayed egg production when infected at the prelay stage of pullets and negatively impacted the laying hens' overall performance.

Host-parasite relationship Octopus tehuelchus – Aggregata valdessensis in Patagonian coast, southwestern Atlantic Ocean

Histopathology associated with the apicomplexan Aggregata valdessensis in the wild population of Octopus tehuelchus was studied. Moreover, to analyze the impact of the prevalence and infection intensity on the dorsal mantle length, sex, maturity stages, condition index and total hemocyte counts on wild O. tehuelchus, Generalized Linear Models (GLMs) were applied. Kruskal–Wallis (KW) test was used to compare growth parameter (RNA/DNA ratios) in infected and uninfected octopuses. Lesions including hypertrophy of host cells and destruction of the tissue organ architecture as well as the replacement of host tissue by parasites were associated with infection. The infection caused an inflammatory defensive response in form of hemocytic infiltration.. The prevalence of apicomplexan increased with octopus dorsal mantle length (GLM, t = 2.68; p = 0.007). Infection intensity was significantly higher in male octopus (GLM, t = 2.69; p = 0.009) and decreased at higher temperatures in both sexes (GLM, t = -2.42; p = 0.018). The digestive gland condition index did not significantly vary with infection intensity, and no significant relationship between total hemocyte counts and coccidian infection was found (Lineal Model, t = 0.46; p = 0.64). The acid nucleic ratio was significantly lower in octopuses infected by A. valdessensis (Kruskal Wallis test, H = 4.8; p = 0.02), suggesting a detrimental effect on O. tehuelchus growth caused by the coccidian.

First Record, Distribution and Occurrence of A Protistan Entomopathogen, Adelina mesnili Perez (Coccidia: Adeleidae) in the Indian Meal Moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae) Populations in Türkiye.

Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae) originating from South America is one of the important insect pests that damages storage products and is found on every continent. There is a new interest in using enthomopathogens for microbial control of P. interpunctella as well as other stored product pests. Coccidia as a group of protistan entomopathogens are host specific and their pathogenic effects on the hosts are more pronounced. Although this pathogenic effect results in increased host mortality or higher susceptibility to insecticides, the suppressive potential of coccidia in natural populations has not been adequately studied. In this study, characterization, distribution and occurrence of a coccidian entomopathogen was aim to show its natural suppressing potential in P. interpunctella populations. During the three years (from 2019 to 2021), a total of 3.432 P. interpunctella samples (2.047 dead and 413 living larvae, 932 adults and 40 pupae) were collected from fourteen populations. After macroscopic examination, suspected samples were dissected in Ringer's solution and then prepared wet smears including host fat body were examined for presence of coccidian pathogens under a light microscope at a magnification of 400-1000X. The oocysts of the coccidian were measured and photographed using a microscope with a digital camera and soft imaging system. The pathogen was observed in the fat bodies of the larvae, pupae and adults. Oocysts measured as 29.52±3.32 (25.27- 35.08) μm in diameter and they include 8 sporocysts. Sporocysts measured as 9.11±0.61 (8.90-9.85) μm. Forty-five of 3.432 P. interpunctella larvae, pupae and adults were found to be infected. Coccidian infections have also reached to the levels that can be considered high in some populations, as significant as 29.2%. The infection was observed in the three (21.4%) of the examined fourteen populations. The coccidian entomopathogen presented in this study is the first Adeleid coccidian record from P. interpunctella populations in Türkiye. The detection of Adelina mesnili Perez (Coccidia: Adeleidae) in at least three populations and the infection rate reaching 29.2 percent, confirms that this pathogen has a considerable effect P. interpunctella populations that cannot be underestimated. Our results confirm that the coccidian pathogen is very effective in the larval stage.

Eimeria granulosa causes spots visible through the serous membrane of small intestine in sheep

Coccidiosis, also known as Eimeriosis, is a highly prevalent parasitic disease affecting sheep in nearly all sheep-rearing nations across the globe. Currently, there is a scarcity of literature documenting the specific lesions in sheep resulting from coccidia infection. This study aimed to investigate these characteristic lesions through necropsy, microscopic observation, and molecular biological techniques. As a result, Eimeria granulosa was identified as the causative agent, which induced distinct pathological alterations in the small intestine of lambs as observed during necropsy. Notably, E. granulosa manifested as small scattered petechiae and white spots, visible through the serous membrane of the small intestine, akin to the pathology observed in E. necatrix. Therefore, this study provides valuable insights for the accurate diagnosis of coccidiosis in sheep.

Effects of xylanase, protease, and xylo-oligosaccharides on growth performance, nutrient utilization, short chain fatty acids, and microbiota in Eimeria-challenged broiler chickens fed high fiber diet

A 21-d experiment was conducted to study the effect of xylanase, protease, and xylo-oligosaccharides on growth performance, nutrient utilization, gene expression of nutrient transporters, cecal short-chain fatty acids (SCFA), and cecal microbiota profile of broilers challenged with mixed Eimeria spp. The study utilized 392 zero-d-old male broiler chicks allocated to 8 treatments in a 4 × 2 factorial arrangement, as follows: corn-soybean meal diet with no enzyme (Con); Con plus xylanase alone (XYL); Con plus xylanase combined with protease (XYL + PRO); or Con plus xylo-oligosaccharides (XOS); with or without Eimeria challenge. Diets were based on a high-fiber (100 g/kg soluble fibers and 14 g/kg insoluble fibers) basal diet. At d 15, birds in challenged treatment were gavaged with a solution containing Eimeria maxima, Eimeria acervulina, and Eimeria tenella oocysts. At d 21, birds were sampled. Eimeria depressed (P < 0.01) growth performance and nutrient utilization, whereas supplementation had no effect. There were significant Eimeria × supplementation interactions for the sugar transporters GLUT5 (P = 0.02), SGLT1 (P = 0.01), SGLT4 (P < 0.01), and peptide transporter PepT1 (P < 0.01) in jejunal mucosa. Eimeria challenge increased the expression of GM-CSF2 (P < 0.01) and IL-17 (P = 0.04) but decreased (P = 0.03) IL-1β expression in the cecal tonsil. Eimeria × supplementation interactions for cecal acetate, butyrate, and total SCFA showed that concentrations increased or tended to be greater in the supplemented treatments, but only in non-challenged birds. Birds challenged with Eimeria spp. had higher concentrations of isobutyrate (P < 0.01), isovalerate (P < 0.01), and valerate (P = 0.02) in cecal content. Eimeria challenge significantly (P < 0.01) decreased the microbial richness and diversity, and increased (P < 0.01) the proportion of Anaerostipes butyraticus, Bifidobacterium pseudolongum, and Lactobacillus pontis. In conclusion, Eimeria infection depressed growth performance, nutrient utilization with regulating nutrient transporters. Furthermore, Eimeria challenge shifted the microbial profile and reduced microbial richness and diversity. On the other hand, enzyme supplementation showed limited benefits, which included increased concentrations of SCFA.

Impacts of Eimeria coinfection on growth performance, intestinal health and immune responses of broiler chickens

Coccidiosis caused by Eimeria is one of the most severe chicken diseases and imposes huge economic losses to the poultry industry globally. Multi-Eimeria species coinfections are common with the most prevalent combination being mixtures of Eimeria acervulina and Eimeria tenella. Although detrimental impacts of either E. acervulina or E. tenella on chicken health are well recognized, no information is available regarding their coinfection effects so far. This study was designed to investigate the influence of coinfection with E. acervulina and E. tenella on broiler chickens. 144 one-day-old broiler chickens within each of trials (trial I or II) were divided into four groups, namely, control group (CG), E. acervulina infection group (EAIG), E. tenella infection group (ETIG) and dual (E. acervulina and E. tenella) infection group (DIG). Then, chickens were measured for weight loss, lesion scores, oocyst outputs, histological changes and expressions of pro-inflammatory (interleukin [IL]−6, IL-8 and IL-18), regulatory (IL-10 and IL-22) cytokines and Toll-like receptors (TLR; TLR2 and TLR4) as well as intestinal barrier (mucin 2 [MUC2] and fattey acid-bingding proteins 2 and 6 [FABP2 and FABP6])- and tight junction (TJ; zonula occluden-1 [ZO-1], occludin [OCLN], and claudins 1 and 5 [CLDN1 and CLDN5])-related proteins at 3, 5, 7, 10, 14 and 21 days post-infection, respectively. Our results consistently showed that although ETIG and DIG exhibited a higher level of weight loss and a more amount of oocyst excretion than EAIG, DIG had lighter lesions than EAIG in the early phase because of coinfection with E. tenella. A higher (P < 0.05) ratio of duodenal villous height to crypt depth was also observed in DIG than EAIG. Moreover, histological changes in the duodenum and cecum varied by single and dual Eimeria infections. Expressions of the intestinal barrier- and TJ-related genes of EAIG, ETIG and DIG were significantly (P < 0.05) upregulated but their levels exhibited differential changes among infected chickens. Similarly, the infected chickens showed significant (P < 0.05) inflammatory responses and higher (P < 0.05) expressions of TLRs in the intestines in comparison to CG. These results presented a comprehensive physiological, pathological and immunological characterization of E. acervulina and E. tenella coinfection in broiler chickens and also shed insights into pathogenesis of multi-coccidia coinfections.

Efficacy of aqueous extract of Vernonia amygdalina leaf against strongyle and coccidia infections in sheep

Gastrointestinal parasites are developing resistance to various commercial anthelmintics. Hence, the need to explore the efficacies of herbal plants against gastrointestinal parasites. The study was therefore conducted to determine the efficacy of aqueous extract of Vernonia amygdalina leaf against strongyles and coccidia spp. in sheep. A total of 60 sheep were used for the study. The animals were randomly assigned to one of the four groups. Those in group A, B, C, and D were given albendazole (ABZ), 10% aqueous V. amygdalina extract (BL10) , 20% aqueous V. amygdalina extract (BL20), and 10 mL distilled water, respectively. All four treatments were given orally, depending on the body weight of the animals, with the exception of the distilled water, which was constant (10 mL) for each animal. Faecal samples were collected from each sheep and examined using McMaster technique. The data were analyzed using R version 4.2. The study brought to light that the prevalence of strongyles in sheep was 70% in the study area at pre-treatment. BL10 had efficacies of 52.58% and 65.08% against strongyles and coccidia spp., respectively. BL20 produced similar anthelmintic effect against strongyles as ABZ. Strongyle spp. showed resistance against ABZ in the study area.

PREVALENCE OF INTESTINAL PROTOZOA, HELMINTHES, AND COCCIDIAN INFECTIONS AMONG PRIMARY SCHOOL CHILDREN IN THALA’A DISTRICT AT AMRAN GOVERNORATE, YEMEN

Background and objectives: The prevalence of intestinal parasite infection, which can range between 18% and 90%, is one of the most frequently reported diseases in Yemen and poses a serious health issue for schoolchildren. In Thala’a District, Amran Governorate, Yemen, the study's main goal was to ascertain the prevalence of intestinal parasite infection among primary school students. Subjects and methods: A cross-sectional survey was conducted from January to April 2022 among 400 schoolchildren, in the age groups 7-12 years. All the children were residents of the city of Thala’a. A pre-tested structured questionnaire was used to collect data. Stool samples were examined for intestinal parasites according to standard laboratory methods. Results: The children's ages ranged from 7 to 12 years, with a mean±SD of 9.52 ± 2.9 years. The results showed that 136/400 (34%) of the tested children were positive for intestinal parasite infection with a higher prevalence of specific intestinal worms (50.4%) than protozoa (33.3%) and coccidian infection (16.3%). The rate of infection alone was also higher than that of multiple parasitic infections (19.3% vs. 14.8%). The highest number of intestinal parasites was 50/400 (12.5%) for Ascaris lumbricoides, and 49/400 (12.25%) for Entameba histolytica followed by 9.8% for Giardia lamblia, 11.5% for Entrobius vermicularis, 6.5% for Cryptosporidium species, 6.5% for Hymenolepis nana, 2% for Schistosoma mansoni, and 0.8% for Trichuris trichiura. Considering the associated factors, boys had a higher infection rate than girls (50% vs. 24.2%) with an OR of 2.6, CI =1.7–3.9 (p &lt; 0.001). A higher rate was also recorded in older children (41%) with an odds ratio of 1.6 (p=0.03). Conclusion: There is a high prevalence of intestinal parasites among school children in Thala’a city, and various control measures are required to control and prevent intestinal parasites among school children. Peer Review History: Received: 3 March 2023; Revised: 22 April; Accepted: 27 June 2023, Available online: 15 July 2023 Academic Editor: Dr. Asia Selman Abdullah, Pharmacy institute, University of Basrah, Iraq, asia_abdullah65@yahoo.com Received file: Reviewer's Comments: Average Peer review marks at initial stage: 5.5/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Rola Jadallah, Arab American University, Palestine, rola@aauj.edu Dr. Tamer Elhabibi, Suez Canal University, Egypt, tamer_hassan@pharm.suez.edu.eg Dr. Sheikh Abdul Khaliq, Department of Pharmacy Practice, Faculty of Pharmacy, Hamdard University, Karachi, Pakistan, drwadhsheikh1974@gmail.com

In vitro and in vivo evaluation of chlorhexidine salts as potential alternatives to potassium dichromate for Eimeria maxima M6 oocyst preservation.

Coccidiosis caused by the Eimeria spp., an Apicomplexan protozoon, is a major intestinal disease that affects the poultry industry. Although most cases of coccidiosis are subclinical, Eimeria infections impair bird health and decrease overall performance, which can result in compromised welfare and major economic losses. Viable sporulated Eimeria oocysts are required for challenge studies and live coccidiosis vaccines. Potassium dichromate (PDC) is typically used as a preservative for these stocks during storage. Although effective and inexpensive, PDC is also toxic and carcinogenic. Chlorhexidine (CHX) salts may be a possible alternative, as this is a widely used disinfectant with less toxicity and no known carcinogenic associations. In vitro testing of CHX gluconate and CHX digluconate exhibited comparable oocyst integrity and viability maintenance with equivalent bacteriostatic and bactericidal activity to PDC. Subsequent use of CHX gluconate or digluconate-preserved Eimeria oocysts, cold-stored at 4°C for 5 months, as the inoculum also resulted in similar oocyst shedding and recovery rates when compared to PDC-preserved oocysts. These data show that using 0.20% CHX gluconate could be a suitable replacement for PDC. Additionally, autofluorescence was used as a method to evaluate oocyst viability. Administration of artificially aged oocysts exhibiting >99% autofluorescence from each preserved treatment resulted in no oocyst output for CHX salt groups.

Effects of Different Eimeria Inoculation Doses on Growth Performance, Daily Feed Intake, Gut Health, Gut Microbiota, Foot Pad Dermatitis, and Eimeria Gene Expression in Broilers Raised in Floor Pens for 35 Days.

The study was conducted to investigate the effects of different Eimeria inoculation doses on the growth performance, gut ecosystem, and body composition of broilers in floor pens for 35 days. A total of 750 15-day-old broilers were allocated to five experimental groups with six replicate pens. The five experimental groups included unchallenged control (CON); Eimeria dose 1 (ED1): E. acervulina: 31,250/E. maxima: 6250/E. tenella: 6250; Eimeria dose 2 (ED2): E. acervulina: 62,500/E. maxima: 12,500/E. tenella: 12,500; Eimeria dose 3 (ED3): E. acervulina: 125,000/E. maxima: 25,000/E. tenella: 25,000; and Eimeria dose 4 (ED4): E. acervulina: 250,000/E. maxima: 50,000/E. tenella: 50,000. On D 21, BW were linearly reduced by increased Eimeria inoculation doses (p < 0.01). On D 35, the Eimeria challenge groups had significantly lower BW compared to the CON group. Increased Eimeria inoculation doses linearly decreased crude fat (CF) (p < 0.01) on D 21. Increased Eimeria inoculation doses tended to increase the relative abundance of the phylum Proteobacteria (p = 0.098) on D 21. On D 35, lean:fat was linearly reduced by increased Eimeria inoculation doses (p < 0.05). Eimeria infection negatively influenced growth performance and gut health in broilers in the acute phase, and the negative effects were prolonged to D 35 in floor pen conditions.

Knowledge, Attitudes and Practices Among Adult Malaria Patients Co-Infected with Opportunistic Intestinal Coccidian Parasites in Fundong Health District, Northwest, Cameroon: A Cross Section Study Design

Introduction: Malaria and intestinal coccidian parasitic co-infections, are becoming a public health emergency affecting millions of people around the world. They are among the leading cause of socio-economic problems, long suffering and death especially in developing countries like Cameroon. Introducing new appropriate preventive and control measures to the population requires thorough assessment of community and patient-based knowledge, attitude and preventive practices towards malaria and pathogenic intestinal coccidian parasites is crucial. Our study aimed to describe knowledge, attitudes, and practices and the risk factors among adult malaria patients co-infected with intestinal parasitic infections in the Fundong Health District, a locality in the Northwest Region of Cameroon. Methods: This was a cross-sectional study carried out between February and December, 2022 involving sixteen (16) health facilities in Fundong Health district. A total number of 330 patients participated in the study. Normal saline wet mount and formol-ether concentration techniques were employed in coccidian parasitic detection. Blood samples were subjected to Giemsa stained and viewed microscopically to detect the Plasmodium parasites. Pretested structured questionnaires were administered to collect information on patient’s socio-demographic factors and determine patients’ knowledge, attitudes and practices towards malaria and intestinal coccidian parasites, as well as their prevention and control. The Pearson’s Chi-Square (χ2) and Student T-test were performed a part of the statistical analysis to check for associations between malaria, infection and between malaria-coccidian co-infection and demographic factors. Statistical significance was set a P-value&lt;0.05. Results: Participants main sources of information on malaria were: Television (TV) and radio 111/330 (34%), community health workers 109/330 (33%), and Hospitals (37%). Participants had good knowledge of mosquito bite as the malaria transmission route, 296/330 (90%), night time is the frequent biting time for the mosquito 296/330(90%), dark corners 229 (69%) and dirty areas 175(53%) as mosquito resting places. Knowledge score was also good on cleaning of the house surroundings, 281/330(85%), clearing the bushes 180/330(55%), poor knowledge score on drainage of stagnant water113/330 (34%) as ways to prevent the mosquito from multiplying. Participants had good knowledge of insecticide treated bed nets 295/330 (89%) as the best way to prevent mosquito bites. However, participants also expressed poor knowledge on edges of the river or streams 7(20%), and animal shed 20/330 (6%) as mosquito hiding places, and Plasmodium specie as malaria causative agent 8/330 (2%) (P-value=0.011). Poor scores were recorded on knowledge of opportunistic intestinal coccidian parasites (19.4%) (P=0.427), and was significantly low on unsafe water (47.2%) (P-value=0.036) as possible transmission routes for coccidian parasites. The knowledge score was also low on the at-risk population for malaria and intestinal coccidian infection (31.9%) P-value=0.009. Participants who had poor knowledge about coccidian transmission routes were less likely to drink from protected water source with an odds of 0.713(95% CI: 0.297-1.711) P value=0.449, compared to those who were more knowledgeable and who were 2.981(95% CI: 1.367-6.115) P value=0.005, more likely to use protected source of drinking water. Participants who had hand washing facility in the household were 3.488 (1.760-6.912) value=0.001 times more likely have better knowledge of coccidian transmission routes compared to those who did not have 0.748 (95% CI: 0.406-1.376) P value=0.350. Poor knowledge score of poor hygiene as major cause of coccidian infection significantly associated with poor practice score of sometime or not at all disinfecting animal shed to prevent coccidian infection among study participants (P =0.039). Conclusion: The overall knowledge scores, attitude and practices level of participants towards malaria and opportunistic intestinal coccidian parasites were relatively good. A significant proportion of the participants still have misconceptions about cause, modes of transmission and practices towards malaria and coccidian prevention methods. A combined health education programmes for malaria and intestinal coccidian parasites aimed at raising community awareness needs to be evidence based and requires innovative approaches, to address the gaps identified in the study.

Graded levels of Eimeria challenge altered the microstructural architecture and reduced the cortical bone growth of femur of Hy-Line W-36 pullets at early stage of growth (0–6 wk of age)

An experiment was carried out to evaluate the impact of mixed Eimeria challenge on skeletal health of Hy-Line W-36 pullets. A total of 540, 16-day-old pullets were randomly allocated into 5 treatment groups, including a nonchallenged control. A mixed Eimeria species solution containing 50,000E. maxima, 50,000E. tenella, and 250,000E. acervulinaoocysts per mL was prepared and challenged to 1 group as a high-dose treatment. The 2-fold serial dilution was done to prepare the medium-high (25,000E. maxima; 25,000E. tenella; 125,000E. acervulina), the medium-low (12,500E. maxima; 12,500E. tenella; 62,500E. acervulina), and the low (6,250E. maxima; 6,250E. tenella; 31,250E. acervulina) dose treatments which were challenged to 3 corresponding groups, respectively. The mineral apposition rate (MAR) was measured from 0 to 14 d post inoculation (DPI) and 14 to 28 DPI using calcein injection. The microstructural architecture of the femur was analyzed using the Skyscan X-ray microtomography (microCT) on 6, 14, and 28 DPI. The results showed that the MAR decreased linearly with an increase in the challenged dose (P < 0.05) during 0 to 14 DPI. The results of microCT revealed that cortical and total BMD, BMC, bone volume (BV), and bone volume as a fraction of tissue volume (BV/TV) of femur decreased both linearly (P < 0.05). Conversely, the total number of pores increased linearly with an increase in challenge dosages on 6 and 14 DPI. Trabecular BMD, BV, BV/TV, trabecular number, and trabecular thickness decreased linearly with an increase in the challenge dosages (P < 0.05) on 6 DPI. Furthermore, Eimeria infection significantly increased the number of osteoclasts and osteoclastic activity (P=0.001). The result of this study suggests that the mixed Eimeria challenge negatively impacts the quality of skeletal health in a linear or quadratic manner with an increase in the concentration of Eimeria oocysts. The negative impact on long bone development might be due to malabsorption, nutrient deficiency during the infection, along with oxidative stress/inflammation disrupting the balance of osteoblastic and osteoclastic cells and their functions.

Amplicon sequencing allows differential quantification of closely related parasite species: an example from rodent Coccidia (Eimeria)

BackgroundQuantifying infection intensity is a common goal in parasitological studies. We have previously shown that the amount of parasite DNA in faecal samples can be a biologically meaningful measure of infection intensity, even if it does not agree well with complementary counts of transmission stages (oocysts in the case of Coccidia). Parasite DNA can be quantified at relatively high throughput using quantitative polymerase chain reaction (qPCR), but amplification needs a high specificity and does not simultaneously distinguish between parasite species. Counting of amplified sequence variants (ASVs) from high-throughput marker gene sequencing using a relatively universal primer pair has the potential to distinguish between closely related co-infecting taxa and to uncover the community diversity, thus being both more specific and more open-ended.MethodsWe here compare qPCR to the sequencing-based amplification using standard PCR and a microfluidics-based PCR to quantify the unicellular parasite Eimeria in experimentally infected mice. We use multiple amplicons to differentially quantify Eimeria spp. in a natural house mouse population.ResultsWe show that sequencing-based quantification has high accuracy. Using a combination of phylogenetic analysis and the co-occurrence network, we distinguish three Eimeria species in naturally infected mice based on multiple marker regions and genes. We investigate geographical and host-related effects on Eimeria spp. community composition and find, as expected, prevalence to be largely explained by sampling locality (farm). Controlling for this effect, the novel approach allowed us to find body condition of mice to be negatively associated with Eimeria spp. abundance.ConclusionsWe conclude that amplicon sequencing provides the underused potential for species distinction and simultaneous quantification of parasites in faecal material. The method allowed us to detect a negative effect of Eimeria infection on the body condition of mice in the natural environment.Graphical

Prevalence of Avian Coccidiosis and Identification of Eimeria spp in Local Broilers and Chickens in Lafia Modern Market, Nassarawa State, Nigeria

Coccidiosis is an important enteric parasitic protozoan disease of poultry. The disease is usually associated with high economic losses to poultry farmers worldwide. This study was conducted to determine the prevalence of coccidiosis, identify the species of circulating Eimeria species, determine the prevalence of Eimeria in respect to breed and gender and determine the deviation of infected birds from normal range of birds slaughtered in Lafia modern market, Nasarawa State, Nigeria. Out of the 204 examined chickens, 36.3% (74/204) of them harbored different Eimeria species. A total of 32.6% (44/74) male and 43.5 (30/74) female were infected. Statistically, no significant difference p&gt;0.05 (χ2=1.559, p=0.2118) was noted between the male and female. Among the local breed, 40.2% (49/74) was infected and for the broilers, 30.5% (25/74) was positive for coccidial infection. Again, there was no statistically significant difference p&gt;0.05 (χ2=1.324, p=0.25) between the local and broiler breeds. In the attempt made to identify the prevalent species of coccidian in the study area, five Eimerian species were identified, namely; Eimeria acervulina, Eimeria brunette, Eimeria mitis, Eimeria necatrix and Eimeria tenella with the prevalence of 14.22%, 5.88%, 1.47%, 6.86%, and 7.84% respectively. Results obtained from this study shows that coccidiosis is an important disease of chickens in the study area and further strategy needs to be implemented to reduce the infection rate of coccidiosis.

Comprehensive analysis of gut microbiome and host transcriptome in chickens after Eimeria tenella infection.

Coccidiosis is an intestinal parasitic disease caused by Eimeria protozoa, which endangers the health and growth of animals, and causes huge economic losses to the poultry industry worldwide every year. Studies have shown that poultry gut microbiota plays an important role in preventing the colonization of pathogens and maintaining the health of the host. Coccidia infection also affects host gene expression. However, the underlying potential relationship between gut microbiome and host transcriptome during E. tenella infection in chickens remain unclear. In this study, metagenomic and transcriptome sequencing were applied to identify microbiota and genes in cecal contents and cecal tissues of infected (JS) and control (JC) chickens on day 4.5 postinfection (pi), respectively. First, microbial sequencing results of cecal contents showed that the abundance of Lactobacillus, Roseburia sp. and Faecalibacterium sp decreased significantly after E. tenella infection (P < 0.05), while the abundance of Alistipes and Prevotella pectinovora increased significantly (P < 0.05). Second, transcriptome sequencing results showed that a total of 434 differentially expressed mRNAs were identified, including 196 up-regulated and 238 down-regulated genes. These differentially expressed genes related to inflammation and immunity, such as GAMA, FABP1, F2RL1 and RSAD2, may play an important role in the process of host resistance to coccidia infection. Functional studies showed that the enriched pathways of differentially expressed genes included the TGF-beta signaling pathway and the ErbB signaling pathways. Finally, the integrated analysis of gut microbiome and host transcriptome suggested that Prevotella pectinovora associated with FABP1, Butyricicoccus porcorum and Colidextribacter sp. associated with RSAD2 were involved in the immune response upon E. tenella infection. In conclusion, this study provides valuable information on the microbiota and key immune genes after chicken E. tenella infection, with the aim of providing reference for the impact of coccidia infection on cecal microbiome and host.

Effect of Nutrients and Herbal Products on Immune Status of Broilers, Experimentally Infected with Mixed Eimeria Species

The present study was conducted to seek herbal alternatives and nutritional modulation to solve the problem of coccidiosis, a deadly protozoan disease of broilers. For the purpose, an experiment of 35 days’ duration (days 0-21 pre-infection period and days 22-35 post-infection period) was conducted to test the efficacy of herbal/natural products/dietary nutrients for the management of mixed coccidial infection prevalent under field conditions. For this purpose wheat grass juice, chelated mineral supplement, herbal anticoccidial and seeds of Embelia ribes without and with sodium bicarbonate, organic chromium, herbal vitamin E-Selenium without and with organic chromium were used to study the immunological parameters of un-infected and infected broilers with 60,000 sporulated oocysts of mixed Eimeria species. Results of the experiment revealed that LST in terms of delta optical density of T- and B-cells was maximum in wheat grass juice while ELISA titre against Newcastle Disease was maximum in herbal vitamin E-Selenium group. All the types of dietary modulation were found to be effective up to different extent in the management of poultry coccidiosis. Further research should focus on micro-array and proteomic technologies to solve the problem.

The effect of dietary Yucca schidigera extract supplementation on productive performance, egg quality, and gut health in laying hens with Clostridium perfringens and coccidia challenge

Yucca schidigera extract (YSE) is a green feed additive that is known to reduce toxic gas emissions and promote intestinal health in animal production. This study investigated the potential of dietary YSE supplementation to mitigate the negative effect of Clostridium perfringens and coccidia infection on productive performance and gut health in laying hens. A total of 48 Lohmann gray laying hens (35 wk of age) were randomly allotted to 1 of 2 groups (n=24) fed with either a basal diet or a YSE-supplemented diet for 45 d. From d 36 to 45, half of the hens in each group were orally administrated with Clostridium perfringens type A and coccidia. This challenge impaired productive performance and egg quality (P < 0.05), destroyed jejunal morphology and functions (P < 0.05), induced jejunal epithelial cell apoptosis (P < 0.05), and downregulated the antioxidant capacity and Nrf2 pathway expression of jejunal mucosa (P < 0.05) in laying hens. Supplementing YSE in the laying hen diet, to some extents, improved productive performance and egg quality (P < 0.05), and alleviated the effect of challenge on morphology, functions, cell apoptosis, and antioxidant capacity in the jejunum (P < 0.05). Overall, the results suggested that dietary YSE supplementation might mitigate the negative effects of Clostridium perfringens and coccidia infection on gut health, and thereby improve the productive performance and egg quality of laying hens, possibly through enhancing the antioxidant capacity of the jejunum.

Faecal prevalence and histopathological evaluation of coccidiosis in bovine calves.

Bovine coccidiois, caused by Eimeria spp. is widely prevalent around the globe and responsible for huge economic losses by causing morbidity and mortality among young calves. The present study was designed to evaluate the prevalence as well as to evaluate histopathological alterations associated with it. The faecal samples were collected from 700 bovine calves upto two month of age from August 2019 to July 2021 and screened for Eimeria oocycts. The intestinal tissue samples of 37 calves were also collected which died during the study period after showing symptoms of diarrhea and examined for histological lesions. The faecal prevalence of Eimeria observed in our study was 2.29% (16/700) while in tissue samples only two out of 37 were found positive for Eimeria infection. Tissue sections revealed various stages of Eimeria gametogony, variable congestion, haemorrhage, and necrosis along with cryptic dilatation and mononuclear cell infiltration. Coccidia was not found to be associated with season, age and sex of calf. Bovine coccidiosis was found to be endemic with low prevalence but severe onset characterized by moderate to severe congestion and inflammatory reaction mainly in the ileum and caecum.