Oocyst Counts Research Articles

Deep glance on the antiparasitic anticancer activities of wheat germ oil in chronically infected immunosuppressed mice with cryptosporidiosis.

Cryptosporidium species are the major cause of water-borne epidemics of diarrhea in both developing and developed countries that vary from self-limited in immunocompetent patients to severe life-threatening in the immunocompromised hosts. There was a proven correlation between cryptosporidiosis and colorectal cancers, although, studies in this field are still limited. Wheat germ oil (WGO) is a natural product with a known antiparasitic effect and potential antiproliferative activities. This study aimed to evaluate the antiparasitic and anticancer activities of WGO in chronically infected immunosuppressed mice compared to Nitazoxanide (NTZ). This experimental case-control study was performed in the period from January till September 2021. Eighty immunosuppressed bred laboratory mice were divided into 4 groups, 20 mice each; GI non-infected; negative control(NC), GII infected non treated; positive control (PC), GII infected, and treated with NTZ, GIV infected, and treated with WGO. Parasitological, histopathological, and immunohistochemical examinations were performed with estimating the rate of maximal survival for the study groups. Parasitological examination revealed a marked reduction in the mean Cryptosporidium spp. oocyst counts in the stool of GIV compared to PC, and GIII (P-value < 0.001). Histopathological and immunohistochemical examinations showed the best results with GIV which revealed restoration of normal villous pattern, with no dysplasia or malignancy could be detected. GIV showed the best survival rate compared to PC and GIII. WGO is an extremely promising agent that has an excellent therapeutic effect against cryptosporidiosis with the ability to control the tumorigenesis process in the chronically infected immunosuppressed hosts.

Characterization of vaccine-induced immune responses against coccidiosis in broiler chickens

Coccidiosis, caused by Eimeria protozoan species, is an economically important enteric disease of poultry. Although commercial live vaccines are widely used for disease control, the vaccine-induced protective immune mechanisms are poorly characterized. The present study used a commercial broiler vaccine containing a mixture of E. acervulina, E. maxima, and E. tenella. One-day-old chicks were vaccinated by spray followed by a challenge at 21 days of age with a mixture of wild type Eimeria species via oral gavage. Oocyst shedding, immune gene expression and cellular responses in the spleen and cecal tonsils were measured at pre- (days 14 and 21) and post-challenge (days 24, 28 and 35) time points. Results showed that the oocyst counts were significantly reduced in the vaccinated chickens at post-challenge compared to unvaccinated control group. While the vaccinated birds had a significantly increased toll-like receptor (TLR) 21 gene expression at pre-challenge, the transcription of interferon (IFN)γ, Interleukin (IL)-12 and CD40 genes in spleen and cecal tonsils of these birds was significantly higher at post-challenge compared to unvaccinated chickens. Cellular immunophenotyping analysis found that vaccination led to increased frequency of macrophages and activated T cells (CD8+CD44+ and CD4+CD44+) in the spleen and cecal tonsils at post-challenge. Furthermore, in vitro stimulation of chicken macrophages (MQ-NCSU cells) with purified individual species of E. acervulina, E. maxima, and E. tenella showed a significantly increased expression of TLR21, TLR2 and IFNγ genes as well as nitric oxide production. Collectively, these findings suggest that TLR21 and TLR2 may be involved in the immune cell recognition of Eimeria parasites and that the vaccine can induce a robust macrophage activation leading to a T helper-1 dominated protective response at both local and systemic lymphoid tissues.

Enumeration and speciation of coccidia affecting turkeys using flow cytometry method

Enumeration of Eimeria oocysts is a common practice in monitoring coccidiosis in turkeys; however, the conventional method of manual microscopic examination of Eimeria oocysts is time-consuming. Previously, we used flow cytometry ( FCM ) to enumerate and speciate coccidia affecting chickens and here, we extended those findings to turkey coccidia species, E. adenoides and E. meleagrimitis . Using FCM, a commercial vaccine containing these species was used to optimize the scatter-plot parameters, including Forward-(size) and Side-(shape/granularity) scatter Area, Height, and Width patterns for Eimeria . The two Eimeria species populations in the vaccine were accurately phenotyped and the gated populations were then sorted using a Cell sorter instrument to obtain pure oocyst suspensions. The individual Eimeria species identity of sorted oocysts was confirmed by PCR using species-specific primers. A significant ( P = 0.0466) correlation (R = 0.9893) in the total oocyst count between FCM and manual methods were observed. Furthermore, when FCM was employed to analyze farm fecal samples, the close similarities in the oocyst morphologies coupled with organic debris particulate interference prevented a precise separation of these 2 species resulting in a lack of oocyst count ( OPG ) correlation between the 2 methods. The OPG counts by FCM were much lower than the manual method; however, a partial OPG trend between the two methods was observed only at the early timepoint collections during a period of 35 d. Collectively, our findings showed that FCM can be used in the enumeration of turkey Eimeria oocysts with a potential scope for a more precise enumeration and speciation in field samples.

The Prevalence of Intestinal Parasite Infections in Goats from Smallholder Farms in Northern Thailand.

SummaryThe present study determined the prevalence of gastrointestinal (GI) parasites in small ruminants kept in smallholder farms in Phitsanulok, Northern Thailand. A total of 885 goats from 31 smallholder farms were selected randomly between May 2019 to January 2020. Fecal samples were collected to estimate fecal egg counts and oocysts counts using the modified McMaster technique. Furthermore, pooled fecal samples from each of the farms were cultured in order to differentiate third-stage larvae of nematode. Examination of fecal samples revealed that 885 were positive for one or more parasites, giving an overall prevalence of 87.2 %, of which 38.6 % were infected with one and 48.6 % with multiple parasite. Strongyles and Eimeria oocysts were the most prevalent. Haemonchus contortus and Strongyloides papillosus were the predominant species based on the percentage of larvae in fecal cultures. Tapeworm eggs were encountered in 14.2 % of all samples. No significant difference in gastrointestinal parasite prevalence was associated with sex of the host. Infection was significantly (P = 0.009) highest in poor body conditioned goats (72.0 %) as compared to moderate (48.9 %) and good body conditioned (50.0 %) goats. This report on prevalence of GI parasites of small ruminants in smallholder farms in Northern Thailand reveals a high endoparasitic infections that appeared well-adapted to environmental conditions. Further studies on endoparasite control are required to establish the impact of parasitism on productive performance, including monitoring control parasite programs are needed for better health and productivity.

Evaluation of the efficacy of cow colostrum in the treatment and its effect on serum proteomes in calves with cryptosporidiosis

This study aimed to investigate the effect of using high-quality colostrum in addition to paromomycin on the treatment outcomes and serum proteomes of calves naturally affected by cryptosporidiosis. Thirty Holstein calves infected with only Cryptosporidium spp. were divided into three equal groups. Calves in the PC group received paromomycin orally at a dose of 100 mg/kg once daily for 5 days. Calves in PCOL and PBCOL groups received 250 ml colostrum 3 h after feeding twice a day for 3 days. The PBCOL group was also given 6 g of sodium bicarbonate 15 min before colostrum administration. While the fecal scores of all calves were evaluated daily for 10 days from the initiation of the treatment, fecal oocyst counts were determined on the 0, 1, 2, 3, 5, 7, and 10th days. Brix%, total protein (TP), immunoglobulin G (IgG), gamma-glutamyltransferase (GGT) levels, and proteomic analyses were performed on the 0 and 3rd days' sera. Considering pretreatment values, fecal scores (8th, 2nd, and 2nd day), and fecal oocyst counts (10th, 3rd, and 2nd day) improved in a significantly (p < 0.05) shorter time in the colostrum groups than in the control group. By serum proteomic analysis, 99, 93, and 83 proteomes were detected in PC, PCOL, and PBCOL groups, respectively. Although the significant changes in any protein in Group PC were absent, significant changes were observed in Alpha-1B-glycoprotein (A1BG), Zinc transporterZIP11 (S39AB), Cathelicidin-1 (CTHL1), Actin_ cytoplasmic-1 (ACTB), and Apolipoprotein A-IV (APOA4) proteins in Group PCOL and Alpha-1-antiproteinase (A1AT), Serum amyloid A protein (SAA), Actin-cytoplasmic-2 (ACTG), Protein HP-20 homolog (HP20) proteins in Group PBCOL with colostral treatment, which indicated that the use of colostrum had an effect on calf serum proteomes. The more pronounced healing and shorter clinical improvement time in the colostrum groups especially colostrum with sodium bicarbonate revealed that these proteomes have positive effects in the treatment with their systemic and local effects in the intestines.

Dietary tea tree (Melaleuca alternifolia) essential oil as alternative to antibiotics alleviates experimentally induced Eimeria tenella challenge in Japanese quails.

Coccidiosis is one of the most common infectious diseases seen in Japanese quails. The current study was conducted to evaluate the impact of tea tree essential oil (TTEO) on growth performance and intestinal health of quails in response to Eimeria tenella challenge. A total of 250 Japanese quails were divided into five treatments: untreated uninfected (negative control); untreated infected (positive control); infected + Amprolium; infected and 1% TTEO; infected and 2% TTEO. Except negative control, all groups were orally dosed with 5 × 104 sporulated oocysts of E. tenella. The results revealed that supplementation of 1% TTEO and treatment of amprolium improved feed intake, weight gain and feed conversion ratio in infected quails compared to the positive control. Similarly, lesion score and mortality was significantly (p < 0.01) reduced in quails supplemented with 2% TTEO and amprolium treated birds. Moreover, oocysts counts and histological features of caecum in infected birds were reversed in 1% TTEO and amprolium treatment. The histological findings of amprolium and 1% TTEO supplemented quails showed intact intestinal villi with mild sloughed epithelium. In conclusion, 1% TTEO can be safely used to control coccidiosis in Japanese quails as natural effective compound.

Administration of spruce bark (Picea abies) extracts in young lambs exhibits anticoccidial effects but reduces milk intake and body weight gain

BackgroundEimeria spp. are widespread apicomplexan parasites known to cause coccidiosis in livestock, resulting in reduced animal welfare and productivity, particularly in sheep. The treatment options are limited, and there is an emerging development of resistance against registered pharmaceuticals. Spruce bark is rich in plant secondary metabolites (PSM), such as condensed tannins, which are bioactive compounds previously shown to have antiparasitic activity. Here, we examined the anticoccidial properties of bark extract of Norway spruce (Picea abies) against a field isolate of ovine Eimeria spp. by treating Eimeria-infected pre-ruminant lambs with water-extracted bark daily for 12 days. We hypothesised that the bark extract would reduce the faecal oocyst excretion and, consequently, the severity of diarrhoea.ResultsOral administration of spruce bark extract significantly reduced the excretion of Eimeria oocysts in milk-fed lambs post treatment till the end of the trial 22 days post infection. This difference in oocyst excretion between the treated and the untreated infected animals increased with time.Compared to the untreated and the sham-infected control group, the group treated with bark extract had softer faeces and reduced milk intake during the treatment period. After discontinuing the treatment, the treated animals got a more solid and formed faeces compared to that of the untreated control group, and the milk intake increased to the level of the sham-infected, untreated control group. The bark extract treated animals had a lower body weight and a lower mean daily body weight gain throughout the whole duration of the experiment.ConclusionsBark extract from Norway spruce showed marked anticoccidial properties by reducing the faecal oocyst count and associated diarrhoea in young lambs. Simultaneously we experienced detrimental effects of the treatment, displayed as reduced feed intake and daily body weight gain. Therefore, we suggest conducting similar studies with lower bark extract dosage to explore the possibilities of a better trade-off to reduce the negative impact while maintaining the antiparasitic effect.

Effect of dehydrated grapefruit peels on intestinal integrity and Eimeria invasion of caprine epithelial cells in vitro and anticoccidial activity in vivo

Anticoccidial drugs are among the preferred resources to control coccidiosis, but the need to have natural alternatives has prompted research into plant products that improve animal performance and might also reduce Eimeria oocyst shedding while avoiding problems of drug residues. Two studies were conducted to evaluate the effects of dehydrated grapefruit peels (GDP) on Eimeria infection and intestinal health. In the first study, an in vitro culture system was used to evaluate the integrity of caprine epithelial cells infected with Eimeria ninakohlyakimovae and incubated with GDP, as well as the infection rate and schizont development of parasites exposed to GDP. In the second study, the anticoccidial effect of the inclusion of 15% GDP1 or 30% GDP2 in the diet and toltrazuril was evaluated in goat kids naturally infected with Eimeria spp. Parameters assessed included the reduction in oocyst counts, weight gain, fecal consistency, intestinal integrity, and malondialdehyde and nitric oxide (NO) production. In vitro, GDP2 and TTZ not only increased transepithelial electric resistance (TEER) values in infected caprine epithelial cells, but also decreased the mean cell invasion rate of sporozoites of E. ninakohlyakimovae and reduced the number and size of schizonts/mm2 of the intestinal cells culture. In vivo, the GDP2 group showed higher mean daily weight gains, reduced fecal oocyst output and decreased oxidative stress in goat kids. These findings demonstrate the beneficial activities of GDP in the enhancement of growth performance and gut health, and the feasibility of using a 30% inclusion of GDP for the replacement of corn in goat production.

Functionally distinct T-helper cell phenotypes predict resistance to different types of parasites in a wild mammal

The adaptive immune system is critical to an effective response to infection in vertebrates, with T-helper (Th) cells pivotal in orchestrating these responses. In natural populations where co-infections are the norm, different Th responses are likely to play an important role in maintaining host health and fitness, a relationship which remains poorly understood in wild animals. In this study, we characterised variation in functionally distinct Th responses in a wild population of Soay sheep by enumerating cells expressing Th-subset specific transcription factors and quantifying Th-associated cytokines. We tested the prediction that raised Th1 and Th2 responses should predict reduced apicomplexan and helminth parasite burdens, respectively. All measures of Th-associated cytokine production increased with age, while Th17- and regulatory Th-associated cytokine production increased more rapidly with age in males than females. Independent of age, sex, and each other, IL-4 and Gata3 negatively predicted gastro-intestinal nematode faecal egg count, while IFN-γ negatively predicted coccidian faecal oocyst count. Our results provide important support from outside the laboratory that Th1 and Th2 responses predict resistance to different kinds of parasites, and illustrate how harnessing specific reagents and tools from laboratory immunology will illuminate our understanding of host-parasite interactions in the wild.

Recovery rates of Eimeria oocysts from cattle feces by Wisconsin sugar flotation method

The purpose of this study was to know the recovery rates of Eimeria oocysts through the course of flotation in sugar solution by WSFM. A fecal sample was collected from a calf infected with Eimeria bovis. Stage one of this study involved recovering oocysts from surface of sugar solution in a test tube with cover slips replaced at time points of 10, 20, 30, 40, 50, 60, 90 min, 2, 3, 4, 5, 6, 24, 48 hr., and 5 days. Oocysts were continuously detected up to 48 hr, with a linear increase in recovery counts for the first 2 hr and the average total count was 953 per gram. Recovery rates of oocysts were 8.1%, 13.3% and 16.4% at 10, 20 and 30 min, respectively. Stage 2 involved a set of 3 tubes. Cover slips were put at intervals of 10, 20 and 30 min on the first tube, while one cover slip was kept for 20 min on the second tube and one for 30 min on the third tube. No significant differences were detected in oocyst counts at 20 min between the first and second tubes, and at 30 min between the first and third tubes. Stage 3 involved the comparison of recovery rates at 10 min between sugar solutions with specific gravities of 1.200 and 1.266. There was no significant difference in oocyst counts between the two solutions. These results indicated that it takes time to recover all Eimeria oocysts from cattle fecal samples by WSFM, but total oocyst counts per gram can be estimated by counting oocysts detected at any sampling time points, such as 10, 20 or 30 min using one cover slip. No improvement of recovery rates was seen by using a higher concentrated sugar solution. Keywords: WSFM, Oocyst, Cattle

Evaluation of anti-malaria potency of wild and genetically modified Enterobacter cloacae expressing effector proteins in Anopheles stephensi

BackgroundMalaria is one of the most lethal infectious diseases in tropical and subtropical areas of the world. Paratransgenesis using symbiotic bacteria offers a sustainable and environmentally friendly strategy to combat this disease. In the study reported here, we evaluated the disruption of malaria transmission in the Anopheles stephensi-Plasmodium berghei assemblage using the wild-type (WT) and three modified strains of the insect gut bacterium, Enterobacter cloacae.MethodsThe assay was carried out using the E. cloacae dissolvens WT and three engineered strains (expressing green fluorescent protein-defensin (GFP-D), scorpine-HasA (S-HasA) and HasA only, respectively). Cotton wool soaked in a solution of 5% (wt/vol) fructose + red dye (1/50 ml) laced with one of the bacterial strains (1 × 109cells/ml) was placed overnight in cages containing female An. stephensi mosquitoes (age: 3–5 days). Each group of sugar-fed mosquitoes was then starved for 4–6 h, following which time they were allowed to blood-feed on P. berghei–infected mice for 20 min in the dark at 17–20 °C. The blood-fed mosquitoes were kept at 19 ± 1 °C and 80 ± 5% relative humidity, and parasite infection was measured by midgut dissection and oocyst counting 10 days post-infection (dpi).ResultsExposure to both WT and genetically modified E. cloacae dissolvens strains significantly (P < 0.0001) disrupted P. berghei development in the midgut of An. stephensi, in comparison with the control group. The mean parasite inhibition of E. cloacaeWT, E. cloacaeHasA, E. cloacaeS−HasA and E. cloacaeGFP−D was measured as 72, 86, 92.5 and 92.8 respectively.ConclusionsThe WT and modified strains of E. cloacae have the potential to abolish oocyst development by providing a physical barrier or through the excretion of intrinsic effector molecules. These findings reinforce the case for the use of either WT or genetically modified strains of E. cloacae bacteria as a powerful tool to combat malaria.Graphical

DNA-based quantification and counting of transmission stages provides different but complementary parasite load estimates: an example from rodent coccidia (Eimeria)

BackgroundCounting parasite transmission stages in faeces is the classical measurement to quantify “parasite load”. DNA-based quantifications of parasite intensities from faecal samples are relatively novel and often validated against such counts. When microscopic and molecular quantifications do not correlate, it is unclear whether oocyst counts or DNA-based intensity better reflects biologically meaningful concepts. Here, we investigate this issue using the example of Eimeria ferrisi (Coccidia), an intracellular parasite of house mice (Mus musculus).MethodsWe performed an infection experiment of house mice with E. ferrisi, in which the intensity of infection correlates with increased health impact on the host, measured as temporary weight loss during infection. We recorded the number of parasite transmissive stages (oocysts) per gram of faeces (OPG) and, as a DNA-based measurement, the number of Eimeria genome copies per gram of faeces for 10 days post-infection (dpi). We assessed weight loss relative to the day of experimental infection as a proxy of host health and evaluated whether DNA or oocyst counts are better predictors of host health.ResultsAbsolute quantification of Eimeria DNA and oocyst counts showed similar but slightly diverging temporal patterns during 10 dpi. We detected Eimeria DNA earlier than the first appearance of oocysts in faeces. Additionally, Eimeria OPGs within each dpi did not explain parasite DNA intensity. Early dpi were characterized by high DNA intensity with low oocyst counts, while late infections showed the opposite pattern. The intensity of Eimeria DNA was consistently a stronger predictor of either maximal weight loss (1 value per animal during the infection course) or weight loss on each day during the experiment when controlling for between-dpi and between-individual variance.ConclusionsEimeria ferrisi oocyst counts correlate weakly with parasite intensity assessed through DNA quantification. DNA is likely partially derived from life-cycle stages other than transmissive oocysts. DNA-based intensities predict health outcomes of infection for the host more robustly than counts of transmissive stages. We conclude that DNA-based quantifications should not necessarily require validation against counts of transmissive stages. Instead, DNA-based load estimates should be evaluated as complementary sources of information with potential specific biological relevance for each host-parasite system.Graphical

Longitudinal dynamics of co-infecting gastrointestinal parasites in a wild sheep population.

Within-year variation in infection is a ubiquitous feature of natural populations, but is determined by a complex interplay of environmental, parasitological and host factors. At the same time, co-infection is the norm in the wild. Longitudinal dynamics of co-infecting parasites may therefore be further complicated by covariation across multiple parasites. Here, we used fecal parasite egg and oocyst counts collected repeatedly from individually marked wild Soay sheep to investigate seasonal dynamics of six gastrointestinal parasite groups. Prevalence and abundance tended to be higher in spring and summer, and abundance was higher in lambs compared to adults. We found that within-year variation in highly prevalent strongyle nematode counts was dependent on adult reproductive status, where reproductive ewes had distinct dynamics compared to males and barren ewes. For similarly prevalent coccidia we found an overall peak in oocyst counts in spring but no differences among males, barren and pregnant ewes. Using multivariate mixed-effects models, we further show that apparent positive correlation between strongyle and coccidia counts was driven by short-term within-individual changes in both counts rather than long-term among-individual covariation. Overall, these results demonstrate that seasonality varies across demographic and parasite groups and highlight the value of investigating co-infection dynamics over time.

S-Methylcysteine Ameliorates the Intestinal Damage Induced by Eimeria tenella Infection via Targeting Oxidative Stress and Inflammatory Modulators.

Avian coccidiosis is one of the major parasitic diseases in the poultry industry. The infection is caused by Eimeria species, and its treatment relies mainly on the administration of anticoccidial drugs, which can result in drug resistance and side effects. The recent trends in avian coccidiosis treatment is directed to the development of a new therapy using herbal compounds. S-Methylcysteine (SMC) is considered one of the organosulfur compounds in garlic that showed promising activity in the treatment of different pathological conditions via a wide range of anti-inflammatory and antioxidant mechanisms. In this study, the anticoccidial activity of SMC was investigated in Eimeria tenella-infected chickens compared to diclazuril as a widely used anticoccidial drug. In this regard, 14-day-old broilers were divided into six groups (n = 18). The first group (G1) was the healthy control group, while the second group (G2) was the non-infected SMC group treated at a dose of 50 mg/kg b.w. (high dose). Moreover, the third group (G3) was the positive control group (infected and non-treated). The fourth group (G4) was the infected group treated with SMC of 25 mg/kg b.w. (low dose), while the fifth group (G5) was the infected group treated with SMC of 50 mg/kg b.w. (high dose). Conversely, the sixth group (G6) was the diclazuril-treated group. The anticoccidial effects of SMC and diclazuril were evaluated by counting oocysts and recording the body weight gain, feed conversion ratio, clinical signs, lesions, and mortality rate. Interestingly, SMC showed potent anticoccidial activity, which was exemplified by reduction of oocyst count. Furthermore, the biochemical, antioxidant, and anti-inflammatory parameters in the cecal tissues were restored toward their control levels in G4, G5, and G6. Histopathological observation of cecal tissues was consistent with the aforementioned results revealing the ameliorative effect of SMC against E. tenella infection. This study concluded novel findings in relation to the anticoccidial role of SMC as a plant-based compound against the E. tenella-induced coccidiosis in broiler chickens combined with its antioxidative and anti-inflammatory properties. Further studies for exploring the mechanistic pathways involved in this activity and the potential benefits from its use in association with conventional anticoccidial drugs are warranted.

Use of Zinc Oxide Nanoparticles as Anticoccidial Agents in Broiler Chickens along with Its Impact on Growth Performance, Antioxidant Status and Hematobiochemical Profile.

The impact of zinc oxide nanoparticles (ZnO-NPs) on the pathogenesis of coccidiosis in broiler chickens was tested. A total of 160 1-day-old broiler chicks (Ross 308) were randomly allocated into 4 groups (n = 40). Group 1: unchallenged, unmedicated; Group 2: challenged, unmedicated; Group 3: challenged, supplemented with diclazuril (1 ppm); Group 4: challenged, supplemented with ZnO-NPs (20 ppm). Mixed Eimeria species (E. maxima, E. acervulina, E. mivati, and E. tenella) of a commercial coccidial vaccine (FORTEGRA®) were used to perform the coccidial challenge by 15× of its vaccinal dose on the 14th day of age. Diclazuril and ZnO-NPs supplementation in Group 3 and 4, respectively, reduced the mortality rate due to coccidial challenge to 5.8% compared to 11.9% in Group 2. The growth performance was improved by ZnO-NPs in coccidiosis-infected group (p ≤ 0.05) compared to Group 2 and was comparable to that of Group 3 (p ≥ 0.05). The average oocyst count was lower in Groups 3 and 4 (7.8 × 103 and 14.3 × 103, respectively) than in Group 2 (67 × 103 oocysts). Group 3 had a decreased gross lesion score in duodenum and caecum (p ≤ 0.05) as well as jujenum and ileum (p ≥ 0.05) compared to Group 2; while the average lesion scores of all intestinal parts in Group 4 were significantly decreased (p ≤ 0.05). However, diclazuril was superior to ZnO-NPs in reducing caecal lesion score (p ≤ 0.05). Plasma carotenoids levels were increased by diclazuril (p ≥ 0.05) and ZnO-NPs (p ≤ 0.05) supplementation compared to Group 2. Oxidative stress appeared on the fourth week post-challenge (pc) in Group 2 (p ≤ 0.05) compared to Group 1, while the dietary supplementation with either diclazuril or ZnO-NPs numerically decreased Malondialdhyde (p ≥ 0.05) and statistically increased antioxidant activity (p ≤ 0.05). Both medications significantly improved the PCV%, Hb% and RBCs count on the 6th-day and 4th-week pc (p ≤ 0.05) compared to Group 2, though this improvement was higher significantly in Group 4 than Group 3 on the 6th day pc (p ≤ 0.05). Neither coccidial challenge nor medications had an impact on the total WBCs count as well as organ index, except Bursa of fabricious index that significantly improved by ZnO-NPs on the 4th-week pc compared to Group 2. Coccidial challenge reduced total protein and globulin levels and increased the serum alanine aminotransferase, serum cholesterol, and low-density lipoprotein levels (p ≤ 0.05) compared to Group 1, while those of both medicated groups (Group 3 and 4) were comparable to Group 1 (p ≥ 0.05). In conclusion, ZnO-NPs were found to be as effective as diclazuril against coccidiosis. However, further research is needed to fully comprehend its anticoccidial mechanisms.

The strong influence of management factors on coccidian infections in smallholder pig farms and the first molecular identification of Cystoisospora suis in Myanmar.

A cross-sectional study was conducted to investigate coccidian infection and associated factors in smallholder pigs, and to identify Cystoisospora oocysts by PCR. A total of 500 pig faecal samples from 330 smallholder farms were collected in Nay Pyi Taw, Myanmar. The faecal flotation method was used to identify Eimeria and Cystoisospora species, and oocyst counts per gram (OPG) of faeces were recorded. Oocysts were differentiated after sporulation. Oocyst DNA was subjected to ITS1-targeted Cystoisospora-specific PCR. The overall coccidian oocyst detection rate by microscopic was 89.0% (445/500). Among the studied samples, 74.0% (370/500) and 70.6% (353/500), were found to be positive with Eimeria spp. and Cystoisospora suis oocysts, respectively. The sequences of C. suis detected were 100% identical to those of C. suis reported from Japan, and had 99.5% resemblance to sequences from Australia and China. Weaner pigs showed the significantly highest (p < 0.05) OPG when compared to other age groups. The highest intensity of coccidian infection (p < 0.05) was found in pigs fed local feed, pigs raised on earthen floors and pigs under poor hygienic conditions. Factors such as age, breed, feed type, and housing floors were found to be significantly associated with coccidian infection (p < 0.05). Age, as well as management factors including floor type, feed type, and hygiene practices on the farm, had a strong influence on the occurrence of coccidian infection in pigs. This is the first study in Myanmar on coccidian infection in pigs and molecular detection of C. suis.

Comparative Efficacy of Moringa oleifera Leaves and Diclazuril on Chicken Coccidiosis

This study aimed to investigate the possible anticoccidial efficacy of Moringa oleifera leaves vs. Diclazuril, as well as production performance and antioxidant activities in broilers exposed to experimental coccidiosis. A total of 160 one-day-old broiler chicks (Ross 308) were randomly allocated into four groups (n=40) as follows: Group 1: unchallenged, untreated; Group 2: challenged, untreated; Group 3: challenged, treated with diclazuril 1mg/kg ration; and Group 4: challenged, treated with Moringa oleifera leaves (20g/kg ration). The birds of all groups except those of group1 were challenged with mixed Eimeria species at the day 14th of the experiment. The mortality rates were 14.8 % in Group 2, 8.8 % in Groups 3 and zero % in group 4. A significant improvement of the final body weight and the feed conversion ratio induced by Moringa oleifera group when compared with that obtained by diclazuril group. The average lesion score was decreased in medicated groups with a better effect in the caecum for Group 3, while Group 4 exhibited a much decrease in lesion score in the upper and the lower intestinal parts and in the caecum. The average oocysts count in Groups 3 and 4 was 7.9x103 and 14.5x103, respectively, while Group 2 had 67.1x103 oocysts. Furthermore, both Moringa oleifera leaves and Diclazuril increased plasma carotenoids levels and growth performance, as well as decreased oxidative stress. Moringa oleifera leaves were found to be as effective as diclazuril against coccidiosis. It could be concluded that, Moringa Oleifera could be useful as alternative product for the control of coccidiosis in poultry production. In addition to its growth promoting effect, as well as its antioxidant effects.

Poultry Coccidiosis-A Concurrent Overview on Etiology, Diagnostic Practices, and Preventive Measures

Coccidiosis is a major parasitic disease in the poultry industry, with great economic implications worldwide. It is a ubiquitous protozoan infection caused by several species of the genus <i>Eimeria</i> (host-specific) that colonize and reproduce in the intestine of birds, ultimately altering the health and performance of the flock. At present, several methods are used to diagnose coccidiosis in poultry, including field and laboratory techniques (intestinal lesion scoring, oocyst counting in feces, and biochemical and molecular diagnosis). Traditionally, diagnosed flocks have been treated either by vaccination to improve the active immunity of the birds against coccidiosis or supplementation of prophylactic anticoccidials to ameliorate the deleterious effects of coccidiosis. However, these methods has certain drawbacks such as vaccine-induced coccidiosis, drug resistance, and residual drug accumulation in the host. Consequently, alternative safe anti-coccidial agents, including the use of phytogenic compounds, have been explored for preventing coccidiosis. Here, we provide a simple overview of the literature on poultry coccidiosis by focusing on the etiology, diagnostic practices, and preventive measures.

Detection of Coccidia Oocysts in Litter and Feces of Broilers in a Floor Pen Trial

Counting oocysts in feces or litter is 1 method to monitor infection levels with Eimeria spp. in chickens after experimental infection or in commercial flocks. Counts of oocysts shed in the feces are thought to follow a typical pattern, with clear peaks representing infection cycles while oocyst counts in litter are representative for at least 2 wk before they begin to deteriorate. The objective of the study was to compare oocyst counts in fresh feces and litter of broilers kept in floor pens with fresh pine shavings as litter material for 42 days. The birds were spray-vaccinated against coccidia in the hatchery. Every 2 to 3 days, 7 pens were sampled by collecting fresh feces and litter from at least 3 locations per pen. Oocysts were counted using a McMaster chamber (Vetslides, Park City, Utah). There were significant differences between pens in oocyst counts in feces as well as in litter, and there were no obvious shedding patterns. Overall, the geometric mean of oocyst counts on day 8 and later was 1,300 oocysts per gram (opg) in feces and 2,700 opg in litter. The variability was lower in litter samples, and the number of pens required to find significant differences between groups in a hypothetical experiment was unrealistically high on most days. Investigating individual fecal samples showed large differences within the pens in addition to the differences between the pens.

Supplementing functional amino acids and polyphenols at low dose can restore performance and amino acid digestibility in broilers challenged with coccidiosis

In broiler chickens, commercial intensive practices exacerbate the pressure of gut heath challenges such as coccidiosis. Coccidiosis impairs nutrient digestion, performance, livability of broiler chickens and reduces overall farm profitability. Literature reports that functional amino acids (AA; arginine, threonine, glutamine) and polyphenols could exert beneficial effects in broilers facing this challenge. The current study aimed at evaluating the efficiency of the supplementation of arginine, threonine and glutamine together with grape extract polyphenols (MIX) on performance, AA digestibility and gut morphology in broiler chickens challenged with coccidiosis.Two hundred eighty-eight (288) newly hatched male broiler chicks of commercial strain (Ross 308) were allocated to 3 treatments with 8 replications. For all except one treatment (Uninfected Untreated Control – UUC), birds were artificially challenged with ADVENT® coccidiosis vaccine 10x dose on day 14. Broiler chickens from the two challenged groups received either no supplementation (Infected Untreated Control – IUC) or a mix of arginine, threonine and glutamine together with grape extract polyphenols at a dose of 1 kg/Ton (IUC+MIX) from day 0 to day 35. Performance and livability were followed during the 35 days of the trial. Lesion scoring, duodenum morphology and fecal oocyst count were assessed on day 21. On day 25 nutrient digestibility was measured on a subset of birds. All data were subjected to an ANOVA and differences among means were detected by multiple range tests.During the course of the trial, coccidiosis challenge reduced growth (-3 g/d, P = 0.008) and feed conversion ratio (-3 pts, P = 0.013) without altering feed intake. It also increased total and species related lesion scores and oocyst counts in feces at d 21 (P < 0.0001). The supplementation of the additive mix restored performance to the same level as the UUC treatment but had no effect on lesion scores and fecal oocyst count. Interestingly, the coccidiosis challenge reduced villous height in duodenum (P = 0.016) which translated into lower digestibility of all AA (P < 0.05), except phenylalanine. The addition of the mix mitigated partially the effect of the challenge on digestibility and to a lesser extent on gut morphology. This study reports that the supplementation of a selected combination of functional amino acids and grape extract polyphenols can fully restore performance of coccidiosis-challenged birds with an improvement of AA digestibility. The results of this study suggest that the mode of action of the mix does not rely on a direct effect on lesion scores, oocyst count or gut morphology and still needs to be determined.