Vector-borne pathogens affecting shelter dogs in eastern Crete, Greece.

Canine pathogens transmitted by blood-sucking arthropods are of significant importance for veterinary and, in some cases, human health. However, they are still underestimated and rarely investigated in many parts of the Mediterranean region, mostly due to financial reasons. Therefore, in the present paper, we investigated the occurrence of blood-associated pathogens affecting dogs in Crete, Greece. For this purpose, blood samples from 103 shelter dogs were screened for the pathogens by PCR and serological tests. Overall, samples from 43 dogs scored positive for at least one pathogen (41.8%). In particular, antibodies to Leishmania infantum were detected in 26 dogs (25.2%), and 15 and 11 animals were positive for Hepatozoon canis (14.6%) and Mycoplasma haemocanis (10.7%) by PCR, respectively. Co-infections were recorded in nine animals. Clinical signs indicative of infection (alterations of skin or coat or reduced body condition) were detected in 10 animals, four of which were infected with one pathogen, three with two pathogens. Based on the results obtained, dogs from Crete appear to be frequently exposed to several blood-borne pathogens, including agents of zoonotic concern. Given that some of the pathogens were reported for the first time in this area, results presented in our study should improve the awareness of the local veterinarians and of dog rescue organisations in order to reduce disease burden on stray and owned dogs and to control the spread of canine vector-borne diseases from Greece to non-endemic areas by travelling or exported infected dogs.

The state of art of neutrophil extracellular traps in protozoan and helminthic infections.

Neutrophil extracellular traps (NETs) are DNA fibers associated with histones, enzymes from neutrophil granules and anti-microbial peptides. NETs are released in a process denominated NETosis, which involves sequential steps that culminate with the DNA extrusion. NETosis has been described as a new mechanism of innate immunity related to defense against different pathogens. The initial studies of NETs were carried out with bacteria and fungi, but currently a large variety of microorganisms capable of inducing NETs have been described including protozoan and helminth parasites. Nevertheless, we have little knowledge about how NETosis process is carried out in response to the parasites, and about its implication in the resolution of this kind of disease. In the best case, the NETs entrap and kill parasites in vitro, but in others, immobilize the parasites without affecting their viability. Moreover, insufficient studies on the NETs in animal models of infections that would help to define their role, and the association of NETs with chronic inflammatory pathologies such as those occurring in several parasitic infections have left open the possibility of NETs contributing to pathology instead of protection. In this review, we focus on the reported mechanisms that lead to NET release by protozoan and helminth parasites and the evidence that support the role of NETosis in the resolution or pathogenesis of parasitic diseases.

Occurrence of tissue cyst forming coccidia in Magellanic penguins (Spheniscus magellanicus) rescued on the coast of Brazil.

The main motivation for this study was to determine the occurrence of Toxoplasma gondii, a cosmopolitan widespread zoonotic parasite distribution that can infect a wide variety of mammals and birds, in Magellanic penguins (Spheniscus magellanicus) in Brazil. In recent decades there has been a significant increase in the number of penguins originating from Argentinian and Chilean Patagonia, where these birds are born, that arrive on the Brazilian coast, where many of them are stranded and rescued. Tissue samples were collected from 330 individuals surveyed from 2012-2015 at the Institute for Marine Animal Research and Rehabilitation (IPRAM) located in Cariacica, state of Espirito Santo, Brazil. Serum were collected from 145 animals surveyed in 2015 for the detection of anti-T. gondii antibodies using the Modified Agglutination Test (MAT ≥20) and 18 birds were positive, with titers of 20 (7 birds), 40 (9 birds) and 80 (2 birds). Mouse bioassay for the isolation of T. gondii was performed using tissues from 54 penguins that were also surveyed in 2015, but no isolates were obtained. DNA from tissue samples of 330 individuals was PCR amplified and sequenced to detect tissue cyst forming coccidians by using pan sarcocystids-directed primers (based on 18S rDNA). These samples were from animals surveyed in 2015 and from frozen stocked tissues from animals surveyed in the years 2012 and 2013. The positives were PCR amplified and sequenced with genus Sarcocystis-specific primers (based on internal transcribed spacer 1, RNA polymerase beta subunit coding gene, and cytochrome B coding gene) and with Sarcocystis falcatula/Sarcocystis neurona- specific primers (based on surface antigens SAG2, SAG3 and SAG4). Sixteen (3.0%) of pectoral muscle samples were positive by all the seven molecular markers and all the samples were identical to each other. Organisms close related to Sarcocystis falcatula were confirmed in all cases. This is the first report on molecular detection of infection by S. falcatula-related organisms and the first report of seropositivity for T. gondii in free-living Magellanic penguins in Brazil. Felids and didephid opossums are definitive hosts of T. gondii and S. falcatula, respectively. Where the penguins acquire the infective forms of the parasites shed by the terrestrial mammals remains to be elucidated.

Recombinant anticoccidial vaccines - a cup half full?

Eimeria species parasites can cause the disease coccidiosis, most notably in chickens. The occurrence of coccidiosis is currently controlled through a combination of good husbandry, chemoprophylaxis and/or live parasite vaccination; however, scalable, cost-effective subunit or recombinant vaccines are required. Many antigens have been proposed for use in novel anticoccidial vaccines, supported by the capacity to reduce disease severity or parasite replication, increase body weight gain in the face of challenge or improve feed conversion under experimental conditions, but none has reached commercial development. Nonetheless, the protection against challenge induced by some antigens has been within the lower range described for the ionophores against susceptible isolates or current live vaccines prior to oocyst recycling. With such levels of efficacy it may be that combinations of anticoccidial antigens already described are sufficient for development as novel multi-valent vaccines, pending identification of optimal delivery systems. Selection of the best antigens to be included in such vaccines can be informed by knowledge defining the natural occurrence of specific antigenic diversity, with relevance to the risk of immediate vaccine breakthrough, and the rate at which parasite genomes can evolve new diversity. For Eimeria, such data are now becoming available for antigens such as apical membrane antigen 1 (AMA1) and immune mapped protein 1 (IMP1) and more are anticipated as high-capacity, high-throughput sequencing technologies become increasingly accessible.

Dual RNA-seq reveals no plastic transcriptional response of the coccidian parasite Eimeria falciformis to host immune defenses.

BACKGROUND: Parasites can either respond to differences in immune defenses that exist between individual hosts plastically or, alternatively, follow a genetically canalized ("hard wired") program of infection. Assuming that large-scale functional plasticity would be discernible in the parasite transcriptome we have performed a dual RNA-seq study of the lifecycle of Eimeria falciformis using infected mice with different immune status as models for coccidian infections. RESULTS: We compared parasite and host transcriptomes (dual transcriptome) between naïve and challenge infected mice, as well as between immune competent and immune deficient ones. Mice with different immune competence show transcriptional differences as well as differences in parasite reproduction (oocyst shedding). Broad gene categories represented by differently abundant host genes indicate enrichments for immune reaction and tissue repair functions. More specifically, TGF-beta, EGF, TNF and IL-1 and IL-6 are examples of functional annotations represented differently depending on host immune status. Much in contrast, parasite transcriptomes were neither different between Coccidia isolated from immune competent and immune deficient mice, nor between those harvested from naïve and challenge infected mice. Instead, parasite transcriptomes have distinct profiles early and late in infection, characterized largely by biosynthesis or motility associated functional gene groups, respectively. Extracellular sporozoite and oocyst stages showed distinct transcriptional profiles and sporozoite transcriptomes were found enriched for species specific genes and likely pathogenicity factors. CONCLUSION: We propose that the niche and host-specific parasite E. falciformis uses a genetically canalized program of infection. This program is likely fixed in an evolutionary process rather than employing phenotypic plasticity to interact with its host. This in turn might limit the potential of the parasite to adapt to new host species or niches, forcing it to coevolve with its host.

A serosurvey of selected cystogenic coccidia in Spanish equids: first detection of anti-Besnoitia spp. specific antibodies in Europe.

BACKGROUND: Equine besnoitiosis, caused by Besnoitia bennetti, and equine protozoal myeloencephalitis (EPM), caused by Sarcocystis neurona and Neospora hughesi are relevant equine diseases in the Americas that have been scarcely studied in Europe. Thus, a serosurvey of these cystogenic coccidia was carried out in Southern Spain. A cross-sectional study was performed and serum samples from horses (n = 553), donkeys (n = 85) and mules (n = 83) were included. An in-house enzyme-linked immunosorbent assay (ELISA) was employed to identify a Besnoitia spp. infection and positive results were confirmed by an a posteriori western blot. For Neospora spp. and Sarcocystis spp., infections were detected using in-house ELISAs based on the parasite surface antigens N. hughesi rNhSAG1 and S. neurona rSnSAG2/3/4. Risk factors associated with these protozoan infections were also investigated. RESULTS: Antibodies against Besnoitia spp., Neospora spp. and Sarcocystis spp. infections were detected in 51 (7.1%), 46 (6.4%) and 20 (2.8%) of 721 equids, respectively. The principal risk factors associated with a higher seroprevalence of Besnoitia spp. were the host species (mule or donkey), the absence of shelter and the absence of a rodent control programme. The presence of rodents was the only risk factor for Neospora spp. infection. CONCLUSIONS: This study was the first extensive serosurvey of Besnoitia spp. infection in European equids accomplished by two complementary tests and gives evidence of the presence of specific antibodies in these populations. However, the origin of the infection is still unclear. Further parasite detection and molecular genotyping are needed to identify the causative Besnoitia and Neospora species. Finally, cross-reactions with antibodies directed against other species of Sarcocystis might explain the positive reactions against the S. neurona antigens.

Investigation of the effect of coccidial vaccine challenge on apparent and standardized ileal amino acid digestibility in grower and finisher broilers and its evaluation in 21-day-old broilers.

The effect of coccidial vaccine challenge (CVC, Coccivac(®)B; challenged, CHA; or unchallenged, NCH) on ileal endogenous amino acid (IEAA) losses and standardized ileal AA digestibility (SIAAD) in 21- and 42-day-old broilers fed a corn-soybean meal-dried distillers' grains with solubles-poultry by-product meal-based diet (Expt. 1) and the effect of supplemental AA in ameliorating the effect of CVC (Expt. 2) were evaluated. Expt. 1 was designed as a 2 (d 21 or 42) x 2 (NCH or CHA) factorial arrangement of treatments with 8 replicates in a complete randomized design. The CVC birds were gavaged with 12 x coccidial vaccine on d 15 and 36 and were sampled on 6 d post challenge. SIAAD was determined by correcting apparent ileal AA digestibility for IEAA losses. Feed intake (FI) and BW gain were higher (P ≤ 0.05) in 21-day-old NCH birds compared to the 21-day-old CHA birds while 42-day-old birds had higher FI and BW gain than 21-day-old NCH and CHA birds. Ileal endogenous nitrogen loss was higher (P ≤ 0.05) in 42-day-old CHA birds compared to 42-day-old NCH birds. Apparent ileal AA digestibility in 21-day-old CHA birds was lower (P ≤ 0.05) than for 21-day-old NCH and 42-day-old NCH and CHA birds. SIAAD in 21-day-old CHA birds was lower (indispensable AA = 15.2 and dispensable AA = 17.8%-unit; P ≤ 0.05) than for 21-day-old NCH and 42-day-old NCH and CHA birds. Apparent ileal digestibility and total tract utilization of DM, N, and energy were not different between d 21 NCH and d 42 NCH and CHA birds but higher (P ≤ 0.05) than for d 21 CHA birds. Supplemental AA restored feed efficiency, but not BW gain, to that of the positive control (0.715 vs. 0.737). Results from Expt.1 showed that CVC significantly influenced ileal AA digestibility in 21- but not in 42-day-old birds. Expt. 2 showed that consideration of SIAAD in feed formulation may benefit feed efficiency.

Characterization of an IgG monoclonal antibody targeted to both tissue cyst and sporocyst walls of Toxoplasma gondii.

Toxoplasma gondii infects animals habiting terrestrial and aquatic environments. Its oocysts and tissue cysts are important for the horizontal transmission of this parasite. The oocyst and tissue cyst walls are crucial for the ability of the parasite to persist in the environment or in animal tissues, respectively. However, the composition of these walls is not well understood. We report the generation of monoclonal antibodies directed against wall components using mice immunized with oocyst antigens of T. gondii. One monoclonal antibody (mAb) G1/19 reacted solely with T. gondii sporozoites. The respective antigen had a relative molecular weight (Mr) of 30 kDa. MAb G1/19 failed to react with sporozoites of any other coccidian parasite species tested (Hammondia hammondi, Hammondia heydorni, Cystoisospora felis, Eimeria bovis, Sarcocystis sp.). Another mAb, designated K8/15-15, recognized antigens in sporocyst walls of the parasite and in the walls of in vivo or in vitro produced tissue cysts, as demonstrated by immunofluorescence and immunoblot assays. Antigens of 80 to a high molecular weight protein of about 350 kDa Mr were recognized by this antibody using antigen extracts from sporocysts, and from in vitro or in vivo generated tissue cysts of the parasite. Tissue cyst and sporocyst walls of H. hammondi and H. heydorni, and tissue cysts of Neospora caninum were also recognized by mAb K8/15-15. Sporocyst walls of C. felis also reacted to this mAb. The cyst walls of Sarcocystis sp. and Besnoitia besnoiti were not recognized by mAb K8/15-15. Reactivity by a single mAb against T. gondii antigens in tissue cysts and sporocysts had not been reported previously. MAb K8/15-15 may be a practical tool for the identification of both cysts and sporocysts of the parasite, and may also be potentially employed in proteomic studies on the identification of new components of the cyst and sporocyst walls of T. gondii.

[Vaccines against livestock parasites: expectations and reality]. / Antiparasitäre Vakzinen beim Nutztier: Wunsch und Wirklichkeit.

Parasitic infections in livestock are of major economic importance. However, increasing resistance against antiparasitic drugs, which is particularly prevalent among parasitic helminths and poultry coccidia, might sooner or later call the economic viability of certain livestock branches into question. Thus, there is a need to develop new efficient parasite control tools. In addition to efforts to discover new antiparasitic compounds or to implement targeted selective treatment strategies, development of vaccines would be a future-orientated alternative. The current review elucidates to what extend antiparasitic livestock vaccines are reality or still expectations.

The influence of dietary zinc source and coccidial vaccine exposure on intracellular zinc homeostasis and immune status in broiler chickens.

Coccidia are protozoal parasites which compromise mucosal integrity of the intestine, potentiating poultry morbidity. The host's Zn status influences the course of infection. Therefore, two experiments were designed to determine how supplemental Zn regimens impacted jejunal and caecal immune status and Zn transporter expression. Coccivac®-B was administered weekly at ten times the recommended dose as a mild coccidial challenge (10 CV). Zn was provided through a basal diet, supplemental zinc sulfate (ZnSO4), or a supplemental 1:1 blend of ZnSO4 and Availa®-Zn (Blend). Mucosal jejunum (Expt 1) and caecal tonsils (Expt 2) were evaluated for intracellular Zn concentrations and phagocytic capacity. Messenger expression of Zn transporters ZnT5, ZnT7, Zip9 and Zip13 were investigated to determine Zn trafficking. With 10 CV, phagocytic capacity was decreased in jejunal cells by 2%. In the caecal tonsils, however, phagocytic capacity increased with challenge, with the magnitude of increase being more pronounced with higher dietary Zn (10 CV × Zn interaction; P = 0.04). Intracellular Zn within caecal tonsils was found significantly reduced with 10 CV (27%, P = 0.0001). 10 CV also resulted in an overall increase in the ratio of Zip:ZnT transporters. With the exception of Zip13 transporter expression, dietary Zn source had little impact on any of the measured cellular parameters. Thus, intestinal mucosal tissues had reductions in intracellular free Zn during coccidial challenge, which was coupled with an upregulation of measured Zip transporters. This suggests that under coccidial challenge, intestinal cells attempt to compensate for the drop in intracellular Zn.

Supplemental dietary L-arginine attenuates intestinal mucosal disruption during a coccidial vaccine challenge in broiler chickens.

The present study investigated the effects of dietary arginine (Arg) supplementation on intestinal structure and functionality in broiler chickens subjected to coccidial challenge. The present study was a randomised complete block design employing a 3 × 2 factorial arrangement (n 8) with three dietary concentrations of Arg (11·1, 13·3 and 20·2 g/kg) with or without coccidial vaccine challenge (unchallenged and coccidial challenge). On day 14, birds were orally administered with coccidial vaccine or saline. On day 21, birds were killed to obtain jejunal tissue and mucosal samples for histological, gene expression and mucosal immunity measurements. Within 7 d of the challenge, there was a decrease in body-weight gain and feed intake, and an increase in the feed:gain ratio (P< 0·05). Jejunal inflammation was evidenced by villus damage, crypt dilation and goblet cell depletion. Coccidial challenge increased mucosal secretory IgA concentration and inflammatory gene (iNOS, IL-1ß, IL-8 and MyD88) mRNA expression levels (P< 0·05), as well as reduced jejunal Mucin-2, IgA and IL-1RI mRNA expression levels (P< 0·05). Increasing Arg concentration (1) increased jejunal villus height (P< 0·05) and linearly increased jejunal crypt depth (P< 0·05); (2) quadratically increased mucosal maltase activity (P< 0·05) and linearly decreased mucosal secretory IgG concentration (P< 0·05) within the coccidiosis-challenged groups; and (3) linearly decreased jejunal Toll-like receptor 4 (TLR4) mRNA expression level (P< 0·05) within the coccidiosis-challenged groups. The mRNA expression of mechanistic target of rapamycin (mTOR) complex 1 pathway genes (mTOR and RPS6KB1) and the anti-apoptosis gene Bcl-2 quadratically responded to increasing dietary Arg supplementation (P< 0·05). These results indicate that dietary Arg supplementation attenuates intestinal mucosal disruption in coccidiosis-challenged chickens probably through suppressing TLR4 and activating mTOR complex 1 pathways.

Neutrophil extracellular traps as innate immune reaction against the emerging apicomplexan parasite Besnoitia besnoiti.

Besnoitia besnoiti infection in cattle is an important emerging protozoan disease in Europe causing economic losses and severe clinical signs, such as generalized dermatitis, orchitis, and vulvitis in affected animals. Neutrophil extracellular trap (NET) formation was recently demonstrated as an important effector mechanism of PMN acting against several invading pathogens. In the present study, interactions of bovine PMN with tachyzoites of B. besnoiti were investigated in this respect in vitro. For the demonstration and quantification of NETs, extracellular DNA was stained by Sytox Orange or Pico Green. Fluorescent illustrations as well as scanning electron microscopy analyses (SEM) showed PMN-promoted NET formation rapidly being induced upon contact with B. besnoiti tachyzoites. Co-localization of extracellular DNA with histones, neutrophil elastase (NE) and myeloperoxidase (MPO) in parasite entrapping structures confirmed the classical characteristics of NET. Exposure of PMN to viable, UV attenuated and dead tachyzoites showed a significant induction of NET formation, but even tachyzoite homogenates significantly promoted NETs when compared to negative controls. NETs were abolished by DNase treatment and were reduced after PMN preincubation with NADPH oxidase-, NE- and MPO-inhibitors. Tachyzoite-triggered NET formation led to parasite entrapment as quantitative assays indicated that about one third of tachyzoites were immobilized in NETs. In consequence, tachyzoites were hampered from active invasion of host cells. Thus, transfer of tachyzoites, previously being confronted with PMN, to adequate host cells resulted in significantly reduced infection rates when compared to PMN-free infection controls. To our knowledge, we here report for the first time B. besnoiti-induced NET formation. Our results indicate that PMN-triggered extracellular traps may represent an important effector mechanism of the host early innate immune response against B. besnoiti which may lead to diminishment of initial parasite infection rates during the acute infection phase.

Effects of dietary enzymes on performance and intestinal goblet cell number of broilers exposed to a live coccidia oocyst vaccine.

An experiment was conducted to evaluate the effects of dietary enzymes on performance, tibia ash, and intestinal goblet cells of broilers administered a live coccidia oocyst vaccine (Coccivac B, Schering Plough, Kenilworth, NJ). Cobb 500 straight-run broilers were obtained and one-half of the chicks were sprayed with the live coccidia oocyst vaccine. Chicks were weighed and placed in battery brooders with respect to nonvaccinated or vaccinated group according to dietary treatment. The 8 dietary treatments were a positive control (0.90% Ca and 0.45% available P), a negative control (NC; 0.80% Ca and 0.35% available P), NC + phytase (PHY), NC + protease (PRO), NC + xylanase (XYL), NC + PHY+ PRO, NC + PHY + XYL, and NC + PHY + PRO + XYL. A diet × vaccination interaction (P > 0.05) was not observed for feed intake or BW gain. Feed conversion ratio was improved (P ≤ 0.05) in birds fed NC + PHY + XYL compared with NC. Vaccination reduced (P ≤ 0.05) feed intake and BW gain from d 0 to 18. Tibia ash was reduced (P ≤ 0.05) in the NC and PRO or XYL diets. Vaccination increased goblet cell numbers in the duodenum of birds fed XYL, whereas no differences were found in goblet cell numbers between nonvaccinated and vaccinated birds in other dietary treatments, which resulted in a diet × vaccination interaction (P ≤ 0.05). Protease decreased and NC + PHY+ PRO increased goblet cells in the jejunum at d 7, which resulted in a diet × vaccination interaction (P ≤ 0.05). At d 18, NC + PHY + XYL was the only diet in which vaccination decreased goblet cells in the jejunum, resulting in a diet × vaccination interaction (P ≤ 0.05). The data indicate that NC + PHY + XYL improved the feed conversion ratio in broilers fed corn-soybean meal diets. The vaccination × dietary enzyme interaction altered the number of goblet cells in the small intestine. Dietary enzyme supplementation did not alleviate reductions in growth performance associated with the use of a live coccidia oocyst vaccine.

Carotenoids modulate the effect of coccidian infection on the condition and immune response in moulting house sparrows.

In the present study, we experimentally manipulated coccidian parasitism and dietary carotenoid availability in a fully factorial experiment in male house sparrows (Passer domesticus Linnaeus), and tested whether carotenoid supplementation reduces the cost of parasitism in terms of condition, moult and immune responses. We found that coccidians have a significant but transient negative effect on body mass, which can be reduced if birds have access to carotenoid supplementation in their diet. Experimental manipulation had no significant effect on the moulting parameters of the birds measured following coccidian infestation and during the whole moulting period. Carotenoid supplementation increased the plasma carotenoid concentration in both infested and medicated birds treated with a coccidiostatic drug; however, after two months exposure to parasites, plasma carotenoid concentration increased only in the carotenoid-supplemented and medicated group whereas no difference was observed between the carotenoid-supplemented and infested and non-supplemented groups. On the contrary, coccidian infestation was not affected by carotenoid supplementation. Experimental infestation decreased the antibody response to sheep red blood cells (SRBCs), although no significant effect was observed in the capacity of the birds to respond to a mitogenic challenge with phytohemagglutinin. Within the experimentally infested groups birds with carotenoid-supplemented food tended to have an increased anti-SRBC humoral immune response. The positive correlation between coccidian infestation and the strength of the humoral immune response against SRBCs in the non-supplemented and infested groups indicates that this part of the immune system plays an important role in defence against these parasites.

Enzymes as feed additive to aid in responses against Eimeria species in coccidia-vaccinated broilers fed corn-soybean meal diets with different protein levels.

This research aimed to evaluate the effects of adding a combination of exogenous enzymes to starter diets varying in protein content and fed to broilers vaccinated at day of hatch with live oocysts and then challenged with mixed Eimeria spp. Five hundred four 1-d-old male Cobb-500 chickens were distributed in 72 cages. The design consisted of 12 treatments. Three anticoccidial control programs [ionophore (IO), coccidian vaccine (COV), and coccidia-vaccine + enzymes (COV + EC)] were evaluated under 3 CP levels (19, 21, and 23%), and 3 unmedicated-uninfected (UU) negative controls were included for each one of the protein levels. All chickens except those in unmedicated-uninfected negative controls were infected at 17 d of age with a mixed oral inoculum of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Live performance, lesion scores, oocyst counts, and samples for gut microflora profiles were evaluated 7 d postinfection. Ileal digestibility of amino acids (IDAA) was determined 8 d postinfection. Microbial communities (MC) were analyzed by G + C%, microbial numbers were counted by flow cytometry, and IgA concentrations were measured by ELISA. The lowest CP diets had poorer (P < or = 0.001) BW gain and feed conversion ratio in the preinfection period. Coccidia-vaccinated broilers had lower performance than the ones fed ionophore diets during pre- and postchallenge periods. Intestinal lesion scores were affected (P < or = 0.05) by anticoccidial control programs, but responses changed according to gut section. Feed additives or vaccination had no effect (P > or = 0.05) on IDAA, and diets with 23% CP had the lowest (P < or = 0.001) IDAA. Coccidial infection had no effect on MC numbers in the ileum but reduced MC numbers in ceca and suppressed ileal IgA production. The COV + EC treatment modulated MC during mixed coccidiosis infection but did not significantly improve chicken performance. Results indicated that feed enzymes may be used to modulate the gut microflora of cocci-vaccinated broiler chickens.

Poultry coccidiosis: recent advancements in control measures and vaccine development.

Coccidiosis is recognized as the major parasitic disease of poultry and is caused by the apicomplexan protozoan Eimeria. Coccidiosis seriously impairs the growth and feed utilization of infected animals resulting in loss of productivity. Conventional disease control strategies rely heavily on chemoprophylaxis and, to a certain extent, live vaccines. Combined, these factors inflict tremendous economic losses to the world poultry industry in excess of USD 3 billion annually. Increasing regulations and bans on the use of anticoccidial drugs coupled with the associated costs in developing new drugs and live vaccines increases the need for the development of novel approaches and alternative control strategies for coccidiosis. This paper aims to review the current progress in understanding the host immune response to Eimeria and discuss current and potential strategies being developed for coccidiosis control in poultry.

Detection of Babesia canis rossi, B. canis vogeli, and Hepatozoon canis in dogs in a village of eastern Sudan by using a screening PCR and sequencing methodologies.

Babesia and Hepatozoon infections of dogs in a village of eastern Sudan were analyzed by using a single PCR and sequencing. Among 78 dogs, 5 were infected with Babesia canis rossi and 2 others were infected with B. canis vogeli. Thirty-three dogs were positive for Hepatozoon. Hepatozoon canis was detected by sequence analysis.

Anti-Hepatozoon canis serum antibodies and gamonts in naturally-occurring canine monocytic ehrlichiosis.

The prevalence of IgG antibodies to Hepatozoon canis and the presence of gamonts in the blood and hemolymphatic tissues were studied in dogs with canine monocytic ehrlichiosis (CME) caused by Ehrlichia canis. Both pathogens are transmitted by the tick Rhipicephalus sanguineus. Forty-five out of 69 (65.2%) dogs with CME were seropositive to H. canis by an enzyme-linked immunosorbent assay (ELISA). Intra-neutrophilic gamonts of H. canis were found in 2 out of 69 dogs (2.9%) comprising 4.5% of the seropositive dogs. The present study indicated that the prevalence of antibodies to H. canis was high among dogs with CME in an area where both infections are endemic. However, previous exposure to H. canis was not found as an important contributor to clinical or clinicopathologic abnormalities found in dogs with CME.

Evaluation of immunosuppressants and dietary mechanisms in an experimental disease model for necrotic enteritis.

Clostridium perfringens (CP) is the etiologic agent of necrotic enteritis (NE). Clinical signs of this disease include depression, decreased appetite, diarrhea, and severe necrosis of the intestinal tract. Understanding the disease progression of NE has been difficult due to its complexity and the involvement of multiple factors (dietary components, immunosuppression, and mechanical irritation of the gut) that appear to contribute to this syndrome. In the present investigation, day-of-hatch broilers were fed a 55% wheat diet and randomly assigned to 1 of 8 groups. Treatments included positive control (CP challenge only), commercial coccidia vaccine (CCV), commercial bursal disease vaccine (CBDV), or the combination of CCV and CBDV, and an appropriate negative control for each (vaccinated and not challenged). Challenged treatment groups received 10(7) cfu of CP twice daily. When compared with controls, broilers in each treatment group had increased (P < or = 0.05) lesion scores, with mean scores of 1.05 and 2.05 in the CP and CBDV + CP treatments, respectively. When compared with controls, the incidence of CP increased (P < or = 0.05) in all treatment groups (73 and 100% in the CCV + CP and CBDV + CP treatment groups, respectively). Compared with controls, percentage mortality increased (P < or = 0.05) from 2% to 26 and 34% in the CP and CBDV + CP treatment groups, respectively. Results of this study indicate that the methodology used provides a good model for studying NE.

Hepatozoon griseisciuri in grey squirrels (Sciurus carolinensis): changes of blood leucocyte numbers resulting from infection.

Numbers of leucocytes in squirrels with gametocytes of Hepatozoon in their blood (infected) were compared with animals without gametocytes (uninfected). Typical values for leucocytes/mm3 blood in uninfected squirrels were: leucocytes 5.7 x 10(3), granulocytes 3.4 x 10(3), lymphocytes 2.0 x 10(3) and monocytes 0.3 x 10(3) cells. Infection caused an increase in monocytes, lymphocytes and granulocytes, and there was a significant positive association between parasitaemia level and numbers of both total leucocytes and monocytes. Infected animals had more uninfected monocytes/mm3 blood than did uninfected animals. The proportions of monocytes were more variable over time in infected animals, but no shift between infected and uninfected status was detected. Transfer of serum from infected squirrels to mice resulted in elevated counts of total blood leucocytes, monocytes and granulocytes, but not of lymphocytes, as compared with controls. Serum from squirrels with high parasitaemias had a more marked effect than serum from squirrels with low parasitaemias. Results indicate an infection-related monocytosis, possibly controlled by cytokines, that increases the number of cells available for invasion by gametocytes, thus enhancing the chances of parasite transmission.