A study of the immunomodulatory effects of coconut oil extract in broilers experimentally infected with velogenic Newcastle disease virus.

Objective: This study aims to evaluate the immunomodulatory effects of coconut oil extract (COE) in broilers experimentally infected with velogenic Newcastle disease virus (vNDV). Methods: A total of 150 broiler birds (day-old) were equally divided into five study groups i.e., negative control, positive control, COE-1, COE-2, and COE-3. On day 10, broilers of groups COE-1, COE-2, and COE-3 were supplemented with 1, 2, and 3 ml of COE respectively per liter of drinking water for 15 days. On day 13, 0.1ml/bird (10-5.25 ELD50) of vNDV was inoculated in broilers of positive control, COE-1, COE-2, and COE-3 groups intramuscularly. During this study, growth performance, morbidity, and mortality rates of each study group were recorded. The antibody titer against NDV was determined on days 7, 14, 21, 28, and 35. The levels of IgY and IgM were also determined on the 7th, 14th, and 21st days post-SRBC inoculation. On day 33, avian tuberculin was injected between the 1st and 2nd toes of the left side (intradermally) to measure lymphoproliferative responses. On day 35, the phagocytic activity in the blood was assessed through a carbon clearance assay by injecting carbon black ink into the right-wing vein. The visceral organs having gross lesions were also collected for histopathology. Results: The COE significantly improved the growth performance, and lowered the morbidity and mortality rates of broilers. There was a significant rise in antibody titers against NDV and levels of IgY and IgM antibodies against SRBC in COE-supplemented broilers. The lymphoproliferative response and phagocytic activity were also enhanced. Among COE-supplemented groups, the broilers of the COE-3 group showed a significant increase in growth performance and boosted immune defense. Conclusions: Coconut oil extract has the potential to boost the growth performance and immune status of broilers. It can be used effectively as a feed additive and alternative to antibiotics to prevent the spread of infectious poultry pathogens.

Recalling the reported toxicity assessment of deoxynivalenol, mitigating strategies and its toxicity mechanisms: Comprehensive review.

Mycotoxins frequently contaminate a variety of food items, posing significant concerns for both food safety and public health. The adverse consequences linked to poisoning from these substances encompass symptoms such as vomiting, loss of appetite, diarrhea, the potential for cancer development, impairments to the immune system, disruptions in neuroendocrine function, genetic damage, and, in severe cases, fatality. The deoxynivalenol (DON) raises significant concerns for both food safety and human health, particularly due to its potential harm to vital organs in the body. It is one of the most prevalent fungal contaminants found in edible items used by humans and animals globally. The presence of harmful mycotoxins, including DON, in food has caused widespread worry. Altered versions of DON have arisen as possible risks to the environment and well-being, as they exhibit a greater propensity to revert back to the original mycotoxins. This can result in the buildup of mycotoxins in both animals and humans, underscoring the pressing requirement for additional investigation into the adverse consequences of these modified mycotoxins. Furthermore, due to the lack of sufficient safety data, accurately evaluating the risk posed by modified mycotoxins remains challenging. Our review study delves into conjugated forms of DON, exploring its structure, toxicity, control strategies, and a novel animal model for assessing its toxicity. Various toxicities, such as acute, sub-acute, chronic, and cellular, are proposed as potential mechanisms contributing to the toxicity of conjugated forms of DON. Additionally, the study offers an overview of DON's toxicity mechanisms and discusses its widespread presence worldwide. A thorough exploration of the health risk evaluation associated with conjugated form of DON is also provided in this discussion.

Pathological effects of feeding aflatoxin-contaminated feed on immune status and reproductive performance of juvenile white leghorn males and its mitigation with ∝-tocopherol and Moringa oleifera.

This study was planned to detect the adverse pathological consequences of aflatoxin B1 in White Leghorn (WLH) layer breeder males. Eight-week-old male layer cockerels were separated into six experimental categories: A group was kept as negative control, offered with normal feed only; group B was fed with 400 ppb amount of aflatoxin, while groups F and D fed with normal feed and supplemented with vitamin E 100 ppm and 1% Moringa oleifera, respectively, whereas groups E and C were fed with 400 ppb aflatoxin containing feed and ameliorated with vitamin E 100 ppm and 1% Moringa oleifera, respectively. This study was continued for 2 months and immunologic disorders and reproductive parameters were observed during the trial. To find out immunological status lymphoproliferative response to phytohemagglutinin-P (PHA-P), antibody titers against sheep red blood cells (SRBCs) and carbon clear assay were performed by collecting samples from five birds from each group. The whole data was measured by ANOVA test, and group means were compared by DMR test by using M-Stat C software. Regarding the reproductive status, spermatogenesis, blood testosterone level, testes weight, testes histology, sperm motility, and morphology were negatively affected by aflatoxins, but these deviations positively ameliorated by vitamin E and Moringa. Vitamin E and Moringa found advantageous in boosting the immune status of affected bird. All the immunological parameters including antibody titers against sheed red blood cells, lymphoproliferative response to avian tuberculin and phagocytic potential of macrophages were suppressed by AFB1 however in control, Moringa and vitamin E groups these immunological responses were significantly higher.

Unraveling the combined deleterious effects of ochratoxin A and atrazine upon broiler's health: Toxicopathological, serum biochemical and immunological perspectives.

Atrazine, a herbicide, is used for eradication of broad-leaved herbs in corn crop; and ochratoxins, particularly ochratoxin A (OTA), are major pollutants of poultry diet. Existence of both of these hazardous chemicals as residues is obvious as elucidated by various epidemiological findings. The present study was designed to investigate toxicopathological, serum biochemical and immunological alterations incurred by atrazine alone and/or, in combination with OTA in broilers. For this purpose, one-day old broiler chicks (n = 180) were purchased from a local hatching unit and were fed two levels of atrazine (50 and 150 mg/kg) and one level of OTA (100 µg/kg) in different combinations. Results of this experiment showed a significant reduction in feed intake, body weight gain, relative organ weights, serum total protein, albumin and globulin while there was a significant increase in urea and creatinine levels, decreased antibody response to sheep red blood cells, reduced lymphoproliferative response and phagocytic capacity in groups given OTA and atrazine individually in feed and these effects became more pronounced when atrazine was given in combination with OTA suggesting synergistic effects of both toxicants for each other.

Aflatoxins associated oxidative stress and immunological alterations are mitigated by dietary supplementation of Pichia kudriavzevii in broiler chicks.

Mycotoxins are the secondary metabolites of certain toxigenic fungi which pose severe health stress in humans, animals and poultry. Certain biological agents and components are used to adsorb mycotoxins in poultry industry which provide promising results in this regard. Pichia kudriazevii (PK), a novel yeast, has the ability to enhance the immune status of poultry chicks. The present study was designed to investigate the ameliorative potential of PK against aflatoxins associated immunosuppression and oxidative stress in broiler chicks. 180-one day old broiler chicks were equally divided into six groups and given different combinations of aflatoxins (300 and 600 µg/kg) and PK (1 g/kg). Parameters studied were antibody response to sheep red blood cells, lymphoproliferative response to PHA-P; phagocytic response by carbon clearance assay system, total antioxidant capacity and total oxidant status of chicks. Results of this experiment confirmed the immunomodulation and antioxidant capacity of PK against 300 µg/kg aflatoxin level. However such amelioration was partial when PK was used with 600 µg/kg aflatoxins. Moreover, the exact ratio of aflatoxin: PK to cause such amelioration still needs to be investigated.

Dietary Trichosporon mycotoxinivoron modulates ochratoxin-A induced altered performance, hepatic and renal antioxidant capacity and tissue injury in broiler chickens.

Ochratoxin A (OTA), an important fungal metabolite in foods and feeds has been shown to induce oxidative stress and cellular injuries to human and animal subjects. This study was designed to investigate the mode of action of a biological modifier Trichosporon mycotoxinivorans (TM), against OTA-mediated oxidative stress and tissue toxicity on broiler chickens. The birds were offered diets supplemented with OTA (0.15 and 0.3 mg/kg feed) and/or TM (0.5, 1.0 g/kg) for 42 days of age, and blood and tissue samples were collected to examine the oxidative stress, biochemical and histopathological parameters. Dietary OTA at all the tested levels induced the hepatic and renal tissue injury as indicated by significant decreased total antioxidant capacity in these organs along with significant decreased (p ≤ 0.05) serum concentrations of total proteins and albumin. The serum concentrations of alanine aminotransferase (ALT) and urea were significantly increased, and these observations were further supported by degenerative changes and increased relative weights of liver and kidneys. The dietary supplementation of TM at both tested levels relieved the detrimental impact of 0.15 and 0.3 mg OTA/kg on the studied parameters. The results of the study demonstrated that dietary TM significantly protects broiler chickens by reducing OTA-induced oxidative damage and tissue injury.

Isolation, characterization and in-vitro antigenicity studies of outer membrane proteins (OMPs) of Salmonella gallinarum coated gold nanoparticles (AuNPs).

The development of biomimetic nanoparticles by combining the natural cellular material with synthetic nanoparticles has inspired innovative therapeutic strategies as compared to traditional line of therapeutics. Therefore, the research was conducted to prepare the outer membrane proteins (OMPs) from Salmonella gallinarum and coated on to the surface of synthetic gold nanoparticles (AuNPs). The size of the final product was characterized by Zeta sizer and Zeta potential. The coating of outer membrane proteins onto the gold nanoparticles (AuNPs) was confirmed by transmission electron microscopy (TEM). The sterility, stability and safety of the prepared antigen was confirmed by standard culture media, in biological buffer solution and in-vitro pyrogenic testing, respectively. In-vitro antigenicity of outer membrane protein alone and coated with gold nanoparticles were observed through macrophage phagocytosis assay. On the basis of results, it was anticipated that bacterial outer membrane protein has immunogenic antigenic properties and coating of bacterial outer membrane protein on the surfaces of synthetic gold nanoparticles not only preserved the complex biological characteristics of bacteria but also provided greater immune responses as compared to outer membrane protein alone. So, it was concluded that the coating of bacterial outer membrane proteins on the surface of synthetic gold nanoparticles have synergistic effects to induce the immune responses and promising potential to develop the effective antibacterial vaccine against salmonellosis in poultry birds.

Identification of potent epitopes on hexon capsid protein and their evaluation as vaccine candidates against infections caused by members of Adenoviridae family.

Adenoviruses cause economically important diseases in vertebrates. Effective vaccines against adenoviral diseases are currently lacking. Here, we report a highly conserved epitopic region on hexon proteins of adenoviruses that generate a strong immune response when used as a virus-like-particle (VLP) vaccine, produced by inserting the epitopic region into the core protein of hepatitis B virus. For evaluation of its protective efficacy, the epitopic region from a representative adenovirus, fowl adenovirus serotype 4 (FAdV-4), was tested as a VLP vaccine which conferred 90% protection against challenge with a virulent FAdV-4 isolate in chickens. Importantly, such a high level of protection is not achieved when the epitopic region is employed as a part of a subunit vaccine. As the sequence and the structure of the epitopic region are highly conserved in hexon proteins of adenoviruses, the epitopic region could be employed as a promising VLP vaccine candidate against adenoviral diseases, in general.

The first complete genome sequence and pathogenicity characterization of fowl adenovirus 11 from chickens with inclusion body hepatitis in Pakistan.

Inclusion body hepatitis (IBH), hydropericardium syndrome, and gizzard erosion associated with fowl adenovirus (FAdV) infections are reported globally and resulted in significant poultry industry economic losses. In 2018, severe IBH appeared in Pakistan in a 17-week-old layer flock. Subsequently, a FAdV-11 strain (designated as PKFAd18) was isolated from liver samples and identified based on phylogenetic analyses of the serotype-specific L1 region of the capsid hexon gene. There is no complete genome sequence of the Pakistani FAdV-11. This study successfully sequenced the complete genome of PKFAd18. The full genome of PKFAd18 contains 43 840 base pairs (bp) with a G + C content of 53.9 %, which is comparable to other FAdV serotypes. Similar to other FAdV-11 strains, PKFAd18 has only one fiber, while FAdV-1 and FAdV-4 have two fibers. Notably, PKFAd18 showed unique characteristics compared to other FAdV-11 strains. A natural large genomic deletion (1215 bp) appeared in tandem repeat region two, relative to the ON-NP2 strain. Phylogenetic analyses of the PKFAd18 penton gene showed higher homology with FAdV-9, highlighting potential natural recombination between FAdV-11 and FAdV-9. Moreover, the pathogenicity of PKFAd18 studied in specific-pathogen-free chickens showed that PKFAd18 is capable of inducing severe IBH and could be responsible for IBH in Pakistan. Thus, the first complete genome of FAdV-11 in Pakistan was sequenced in this study, which enriches the diversity of knowledge about FAdV-11 and is useful for developing diagnostics and vaccines for IBH induced by FAdV-11 in Pakistan.

Decanoyl-Arg-Val-Lys-Arg-Chloromethylketone: An Antiviral Compound That Acts against Flaviviruses through the Inhibition of Furin-Mediated prM Cleavage.

Flaviviruses, such as Zika virus (ZIKV), Japanese encephalitis virus (JEV), Dengue virus (DENV), and West Nile virus (WNV), are important arthropod-borne pathogens that present an immense global health problem. Their unpredictable disease severity, unusual clinical features, and severe neurological manifestations underscore an urgent need for antiviral interventions. Furin, a host proprotein convertase, is a key contender in processing flavivirus prM protein to M protein, turning the inert virus to an infectious particle. For this reason, the current study was planned to evaluate the antiviral activity of decanoyl-Arg-Val-Lys-Arg-chloromethylketone, a specific furin inhibitor, against flaviviruses, including ZIKV and JEV. Analysis of viral proteins revealed a significant increase in the prM/E index of ZIKV or JEV in dec-RVKR-cmk-treated Vero cells compared to DMSO-treated control cells, indicating dec-RVKR-cmk inhibits prM cleavage. Plaque assay, qRT-PCR, and immunofluorescence assay revealed a strong antiviral activity of dec-RVKR-cmk against ZIKV and JEV in terms of the reduction in virus progeny titer and in viral RNA and protein production in both mammalian cells and mosquito cells. Time-of-drug addition assay revealed that the maximum reduction of virus titer was observed in post-infection treatment. Furthermore, our results showed that dec-RVKR-cmk exerts its inhibitory action on the virus release and next round infectivity but not on viral RNA replication. Taken together, our study highlights an interesting antiviral activity of dec-RVKR-cmk against flaviviruses.

Potential of botanical driven essential oils against Haemonchus contortus in small ruminants / Potencial de aceites esenciales botánicos contra Haemonchus contortus en pequeños rumiantes

The livestock sector is continuously facing problems in controlling parasitic diseases especially Haemonchosis due to emergence of anthelminthic resistance and failure in vaccination control programmes. Therefore, to increase milk and meat production and emerging demand of meat free from drug residues development of new alternative approaches are appealing for prevention and control of Haemonchosis in small ruminants. Among alternatives, plants driven essentials oils have shown promising results in control of Haemonchus contortus infection at various concentrations by different assays including egg hatch assay, larval development assay, larval exsheathment assay and adult motility assay. Essential oils are complex mixtures of various impulsive or volatile compounds which have potential to control Haemonchosis. The current study reviews the therapeutic effects of essential oils of plants against Haemonchus contortus and to be used them against Haemonchus contortus for future perspectives.

El sector ganadero enfrenta continuamente problemas para controlar las enfermedades parasitarias, especialmente la hemoncosis, debido a la aparición de resistencia antihelmíntica y al fracaso en los programas de control de vacunación. Por lo tanto, para aumentar la producción de leche y carne, y la demanda emergente de carne libre de residuos de medicamentos, el desarrollo de nuevos enfoques alternativos es atractivo para la prevención y el control de la hemoncosis en pequeños rumiantes. Entre las alternativas, los aceites esenciales producidos por las plantas han mostrado resultados prometedores en el control de la infección por Haemonchus contortus a diversas concentraciones mediante diferentes ensayos, incluido el análisis de eclosión de huevos, el desarrollo de larvas, el análisis de vaciado de larvas y el ensayo de motilidad en adultos. Los aceites esenciales son mezclas complejas de varios compuestos impulsivos o volátiles que tienen potencial para controlar la hemonchosis. Este estudio revisa los efectos terapéuticos de los aceites esenciales de las plantas contra Haemonchus contortus y evalúa sus perspectivas futuras como agentes para combatir las enfermedades causadas por este parásito.

Impact of dietary Trichosporon mycotoxinivorans on ochratoxin A induced immunotoxicity; In vivo study.

Ochratoxin A (OA), the secondary metabolite of certain Aspergillus and Penicillium species, is one of the potent biological immune-suppressor. The present study was designed to explore the in-vivo efficacy of Trichosporon mycotoxinivorans (TR); yeast strain isolated from the hindgut of the termite Mastotermes darwiniensis, against the immunotoxicity of OA in broiler birds. For this purpose, broiler chicks were offered diet added with TR (0.5, 1.0 or 2.0 g/kg feed) and/or OA (0.15, 0.3 or 1.0 mg/kg feed) for 42 days. Dietary OA at all levels, resulted in significant reduction (p ≤ 0.05) in the immune response of broiler birds as recorded by vacuolation and darkly stained pyknotic nuclei in bursa of Fabricius and thymus, humoral immune responses to sheep red blood cells (SRBC), in-vivo lymphoproliferative response to Phytohemagglutinin-P (PHA-P) and mononuclear phagocytic system function assay. Addition of TR in broiler diet significantly reduced (p ≤ 0.05) the immunotoxicity of OA at 0.15 and 0.30 mg/kg; however, against higher dietary level of OA (1.0 mg/kg), a partial protection was observed. Feeding TR alone had no immunomodulatory effect at any of tested level. Dietary addition of TR is proposed as an approach to combat the OA mediated immunological damages in broiler birds.

Amelioration of toxicopathological effects of cadmium with silymarin and milk thistle in male Japanese quail (Coturnix japonica).

Cadmium is an important widely distributed heavy metal in the environment due to its several industrial uses, while milk thistle is an important herb and is a source of several antioxidant particularly silymarin which is a pharmacological active substance present in seeds of milk thistle plant (Silybum marianum). The current study investigated pathological effects of cadmium (Cd) and their amelioration with silymarin (SL) and milk thistle (MT) quails. A total of 144 quails were equally divided into 9 groups and given different combinations of cadmium chloride (150 and 300 mg/kg feed), SL (250 mg/kg), and MT (10 g/kg) feed. Parameters studied were clinical signs, mortality, organ weights, testes weight and volume, and gross and microscopic pathology. Results of this study indicated an increased mortality and reduced body weight in cadmium-treated quails. Quails were dull, depressed compared with control. Swollen hemorrhagic liver along with atrophied testes were also observed in these groups. No active spermatozoa were observed in lumen of seminiferous tubules of Cd-treated birds presenting arrest of spermatogenesis. Supplementing MT and SL ameliorated mortality, organ weights, spermatogenesis, and histopathological lesions. It may be concluded that MT and SL proved beneficial in cadmium-induced toxicities in Japanese quails.

Dietary L-carnitine and vitamin-E; a strategy to combat ochratoxin-A induced immunosuppression.

This study aimed to evaluate the effect of dietary ochratoxin A (OA), in the presence and absence of L-carnitine (LC) and vitamin E (VE), on the humoral immune responses of White Leghorn cockerels (WLC). One-day old white male Leghorn chicks were divided into 12 groups, having 20 birds each and were offered ration contaminated with OA (1.0 or 2.0 mg/kg feed) alone and concurrently with LC (1.0 g/kg) and/or VE (0.2 g/kg), for 42 days. The humoral immune responses were accessed by lymphoproliferative response to avian tuberculin, in-vivo phagosomes activity to carbon particles and antibody response to the sheep red blood cells (SRBCs). The dietary addition of OA alone suppressed the humoral immune responses, however, the exposure of birds to 1.0 mg/kg OA in the presence of LC and/or VE showed a significant reduction in OA induced immunotoxicity. This protective response was absent in the birds fed 2.0 mg/kg OA in the presence and absence of LC and/or VE. Histopathological and morphometric examination of the bursa of Fabricius exhibited a decrease in the severity and frequency of OA induced lesions in the presence of dietary LC and/or VE. The use of LC and VE as dietary supplement, can effectively overcome OA (≤1.0 mg/kg) induced immunosuppression.

Acaricidal and insecticidal effects of essential oils against ectoparasites of veterinary importance / Efectos acaricidas e insecticidas de los aceites esenciales contra los ectoparásitos de importancia veterinaria

Ectoparasitism in animals has become an issue of great concern that needs to be resolved to prevent huge economic losses occurring to livestock industry all over the world. Synthetic adrugs have been playing a major role in controlling ectoparasites, but their frequent and irrational use has resulted in drug resistance to routinely used chemicals and their residual effects on food and environment. Therefore, this approach of using chemical acaricides and insecticides is losing its popularity and effectiveness in controlling ectoparasites. So, the development of alternative approaches in ectoparasite management is currently required. Among alternative protocols, plants and their essential oils have played remarkable role in controlling different ectoparasites (ticks, flies, mites, lice) of veterinary importance. Essential oils have been proved to be cheaper, more effective and safer therapeautic agents against different ectoparasites of livestock importance.

En los animales el ectoparasitismo se ha convertido en un tema de gran preocupación que debe resolverse para evitar que se produzcan grandes pérdidas económicas para la industria ganadera en todo el mundo. Los aditivos sintéticos han desempeñado un papel importante en el control de los ectoparásitos, pero su uso frecuente e irracional ha dado como resultado la resistencia a los fármacos utilizados habitualmente y efectos residuales sobre los alimentos y el medio ambiente. Por lo tanto, el enfoque basado en el uso de acaricidas e insecticidas químicos está perdiendo popularidad y efectividad en el control de los ectoparásitos. Por lo tanto, actualmente se requiere el desarrollo de enfoques alternativos en el manejo de ectoparásitos. Entre los protocolos alternativos, las plantas y sus aceites esenciales han jugado un papel notable en el control de diferentes ectoparásitos (garrapatas, moscas, ácaros, piojos) de importancia veterinaria. Se ha demostrado que los aceites esenciales son agentes terapéuticos más baratos, más efectivos y más seguros contra diferentes ectoparásitos de importancia ganadera.

Pathological effects of concurrent administration of aflatoxin B1 and fowl adenovirus-4 in broiler chicks.

The current study was designed to investigate pathological effects of fowl adenovirus in broilers exposed to aflatoxin B1. Fowl Adenovirus-4 (FAdV-4) infection is remerging in all types of poultry birds in Pakistan. Poultry feed contamination with mycotoxin (aflatoxin) is another important global issue. A total of 125-day old broiler birds were divided into six equal groups. Group A served as control. B and C groups were administered with aflatoxin B1 (AFB1) 100 and 200 µg/kg feed. Group D was infected with FAdV-4, while groups E and F administered with both AFB1 (100 & 200 µg/kg) along with FAdV-4. These birds were monitored for clinical signs and mortality. Feed intake, body weight (BW), relative organ weights and gross & histopathological lesions were recorded. The highest mortality was observed in group F (FAdV-4 + AFB1 200 µg/kg feed) and the lowest mortality was observed in group B (AFB1 100 µg/kg feed). Body weights of all the groups were significantly (p < 0.05) lower as compared with control group. Relative weight of liver and kidneys in groups E and F were significantly higher as compared with control. Grossly, liver was swollen, anemic with round margins in groups D, E and F. Kidneys were also swollen with whitish areas indicating dead tissue. Microscopically intranuclear inclusion bodies were observed in group D-F. The hepatic parenchyma was also indicating necrotic changes along with vacuolar degeneration. In renal parenchyma, acute tubular necrosis was observed in groups C, E and F. It was concluded that AFB1 intoxication lead to dose dependent changes in liver and kidneys. Severity of the changes was increased in interactive groups of AFB1 with FAdV-4. Therefore, feed should be regularly monitored for AFB1 levels and day old chicks for vertically transmitted FAdV-4 to prevent losses.

Potential for amelioration of aflatoxin B1-induced immunotoxic effects in progeny of White Leghorn breeder hens co-exposed to vitamin E.

This study was designed to evaluate the protective activity of Vitamin E (Vit E) on the immunotoxic effects induced by aflatoxin B1 (AFB1) in the progeny of breeder hens. For this purpose, 192 White Leghorn (WL) layer breeder hens were divided into 12 groups (A-L) and then fed test diets for either 1, 2 or 3 weeks. Group A was kept on basal feed (2900 Kcal/kg metabolizable energy) and served as control, while group B was offered a feed supplemented with Vit E at 100 mg/Kg. Groups C-G were offered feed containing 0.1, 0.5, 2.5, 5.0, and 10.0 mg/Kg AFB1, respectively, whereas groups H-L were offered the same dietary levels of AFB1 along with 100 mg/Kg Vit E supplementation. Hatching eggs were shifted to an incubator on a weekly basis to get progeny chicks. Hatched chicks in each group were maintained on basal ration and then subjected to different immunological assays. Lymphoproliferative responses (against PHA-P), antibody titers (against SRBC), oxidative damage to RBC, as well as phagocytic and nitrite production potential of the peritoneal macrophages from the chicks, were all adversely impacted by hen exposure to the higher doses of AFB1 or by increased intake (time) by the hens at a given dose of the toxin. No consistent ameliorative effects from Vit E were noted in these studies, i.e. effects seen against lower AFB1 doses were no longer apparent with the highest doses of AFB1. As such, for now it can be concluded that, with this particular single dose level of Vit E, AFB1-associated immunotoxic effects in progeny chicks can potentially be mitigated by dietary intake of Vit E by their hen dams. However, this is clearly an outcome that is driven by the level of the mycotoxin present in the feed. Future studies need to examine what impact higher Vit E doses than those employed herein might have in these ameliorative outcomes.

Amelioration of Ochratoxin A-induced immunotoxic effects by silymarin and Vitamin E in White Leghorn cockerels.

Silymarin (SL) is the bioactive extract of the plant Silybum marianum and Vitamin E (VE) is an important anti-oxidant. The present study was designed to evaluate potential ameliorative effects of SL and VE against Ochratoxin A (OTA)-induced immunotoxic effects in White Leghorn cockerels. One day-old birds were divided into 12 groups (20 birds/group) and fed basal diets amended with OTA (1.0 or 2.0 mg/kg) alone or in combination with SL (10 g/kg) and/or VE (200 mg/kg) for 42 days. Immunological in situ responses, including antibody formation against sheep red blood cells (7 and 14 days after both primary and booster injections), lymphoproliferative responses to avian tuberculin (30 days of age), and mononuclear phagocytic system function (i.e. by clearance of injected carbon particles) assay (42 days of age), were assessed. Results suggested that silymarin and Vitamin E alone or in combination ameliorated the immunotoxic effects induced by 1.0 mg OTA/kg but could not significantly impact on the effect from ingestion of 2.0 mg OTA/kg. The results of the present study suggested that both SL and VE possess an ability to ameliorate OTA-induced immunotoxicity in chicks. However, it remains to be determined whether/what SL:OTA or VE:OTA ratios are required to assure such mitigation of OTA-induced immunotoxicities.

Toxico-pathological effects of in ovo inoculation of ochratoxin A (OTA) in chick embryos and subsequently in hatched chicks.

This study was designed to investigate the toxico-pathological effects of in ovo inoculation of ochratoxin A (OTA) in chicken embryos and subsequently in the hatching chicks. Nine hundred fertile white leghorn (WL) layer breeder eggs were divided into eight groups (A-H). Group A was maintained as untreated control, whereas group B was kept as sham control (10 µL of 0.1 M NaHCO(3) solution). Before incubation, groups C, D, E, F, G, and H were injected with 0.01, 0.03, 0.05, 0.10, 0.50, and 1.00 µg OTA/egg, respectively. At 53 hrs of incubation, crown to rump length, optic cups, and eye lens diameters were significantly (p ≤ .05) lower, whereas neural tube closure defects were higher in the OTA-treated embryos. Teratogenic defects (studied at day 9 of incubation) and embryonic mortalities were higher in the groups administered high doses of OTA. A significant increase was noted in the serum concentration of ALT, urea, and creatinine, along with higher weights of liver and kidney, in chicks hatched from OTA-contaminated eggs. These findings suggested that there are teratogenic and substantive toxicological risks in the developing chicken embryos and hatched chicks that could be exposed to OTA in ovo.

Immunological responses of male White Leghorn chicks kept on ochratoxin A (OTA)-contaminated feed.

This study was designed to evaluate some immunological responses of male White Leghorn (WL) chicks kept on an ochratoxin A (OTA)-contaminated diet. For this purpose, 350 1-day-old male WL chicks were divided into five groups (A-E). Group A was kept as control, while Groups B, C, D, and E were fed OTA-contaminated feed at 0.1, 0.5, 1.0, and 1.5 mg/Kg diet, respectively, for 21 days, and then basal ration for the remaining period. At 14- and 16-days of age, random chicks (n = 10) from each group were used for analyses of phagocytic function of the reticuloendothelial system or for measuring the lymphoproliferative responses to intradermally-administered T-cell mitogen, phytohemagglutinin-P (PHA-P), respectively. At 30-days of age, abdominal macrophages were collected from 15 chicks/group and utilized for determination of their phagocytic potential and for nitrite production. Antibody (Ab) titers (i.e., total antibodies, IgM, and IgG) against sheep red blood cells (SRBC) were determined at 7 and 14 days after a primary (at 7 days of age) and a booster (given 14 days after primary [at 21-days of age]) dose (intravenous) of the antigen. Data from the present study showed that the relative weight of the bursa of Fabricius of chicks fed OTA for 14 and 21 days and the spleen of chicks fed OTA for 21 days were significantly lower than their control counterpart. Phagocytic function of reticuloendothelial system evaluated by carbon clearance, and lymphoproliferative response to PHA-P, of chicks kept on OTA-contaminated diet were significantly lowered. The percentage of abdominal macrophages displaying phagocytosis of SRBC, the number of SRBC/macrophage, and nitrite production were each significantly lower in cells from chicks in the OTA-fed groups. Total Ab (at days 7 and 14 post-booster SRBC injection) and IgG (at day 14 post-primary and day 7 post-booster SRBC injection) titers against SRBC showed significant reductions in the groups fed OTA-contaminated diet. The findings of this study are in line with the previous work suggesting the immunosuppressive effect of OTA in male WL chicks regarding functional impairment in some of the components of the immune system.