Therapeutics effects of bovine colostrum applications on gastrointestinal diseases: a systematic review.

BACKGROUND: Evidence on the effects of bovine colostrum (BC) supplementation on gastrointestinal (GI) diseases is conflicting. OBJECTIVES: This systematic review summarized the findings of clinical trials (CTs) on the effects of BC supplementation on GI diseases. METHODS: A systematic search was conducted in online databases, including PubMed, ISI Web of Science, and Scopus, until March 2021 and updated until December 2023. CTs investigated BC's effect on any measurable symptomatic change in terms of GI health as the primary outcome variable or as one of the outcomes in any population eligible for this systematic review. RESULTS: Out of 6881 records, 22 CTs (uncontrolled = 4, cross-over = 1, and parallel = 17) with 1427 patients were enrolled in the systematic review. Diarrhea, the most frequently evaluated symptom (20 interventional arms), was decreased in frequency with BC supplementation in 15 of these arms. However, most studies reported no change in its duration. BC supplementation consistently reduced stool frequency across all seven studies. Abdominal pain relief was noted in four interventional arms but showed no improvement in five others. Assessment of other GI symptoms was limited, yielding inconclusive results. CONCLUSIONS: There is limited evidence on the effects of BC on GI diseases, with mixed findings. More well-designed controlled clinical trials are required to explore its effects.

Bovine Colostrum in Increased Intestinal Permeability in Healthy Athletes and Patients: A Meta-Analysis of Randomized Clinical Trials.

BACKGROUND: Increasing intestinal permeability causes chronic inflammation, which is one of the etiological factors of many diseases that presently constitute global challenges. AIMS: Considering the importance of developing therapies to eliminate the increased intestinal permeability, in this systematic review and meta-analysis, we analyze the impact of bovine colostrum (BC) on the gut barrier and its permeability. METHODS: Online databases, including PubMed, ISI Web of Science, and Scopus, were searched to find pertinent articles up to March 2022. Weighted mean difference (WMD) and 95% confidence intervals (CI) were considered as effect sizes. The random-effects model was used to pool the study results. RESULTS: A total of ten articles were included in the meta-analysis. The pooled effect revealed a significant reduction in the 5-h urinary lactulose/rhamnose ratio after BC consumption [mean difference (MD): -0.24; 95% CI -0.43 to -0.04; I2 = 99%] and urinary lactulose/mannitol ratio (MD: -0.01; 95% CI -0.02 to -0.001; I2 = 29.8%). No differences were observed in the plasma intestinal fatty acid-binding protein (I-FABP) between BC and control groups (MD: 2.30; 95% CI -293.9 to 298.5; I2 = 92%). CONCLUSIONS: BC supplementation significantly reduced intestinal permeability; however, to confirm the results, more randomized clinical trials considering different quality, dose, and duration are needed.

Apoptosis in field and experimental cases of avian influenza H9N2 infection in broiler chickens in Iran.

Avian influenza virus subtype H9N2 is widely circulating around the globe affecting many species of animal including mammals and birds as well as human beings. The virus has pandemic potential due to segmented nature of the viral genome. Ultra-structural features of apoptosis in field and experimental infection of H9N2 avian influenza virus were studied. Freshly dead birds from affected broiler farms and experimentally infected broiler chickens with H9N2 subtypes were subjected to routine necropsy. Post-mortem findings in different organs were recorded. Appropriate specimens from the trachea were taken for electron microscopy studies. In electron microscopy study, frequent apoptotic bodies were observed in the epithelial cells of the trachea. Increase of antibody titer to H9N2 virus following challenge with the virus in experimental group indicates that the infectious cycle has been initiated in the affected birds.

Hyper-Immune Bovine Milk as an Immunological and Nutritional Supplement for COVID-19.

Many different strategies have been used to fight against the Coronavirus disease (COVID-19) pandemic as a therapeutics or prophylaxis approaches. However, not enough attention has been paid to general and specific immune factors and nutritional components found in hyper-immunized dairy products. Hyper-immune bovine colostrum (HBC) has been used against many different respiratory and gastrointestinal tracts infections during past decades. An isolated dairy farm was established, and nine mixed Holstein X Simmental dairy cattle in their 6-7 months of gestation period were chosen for hyper-immunization with inactivated Severe acute respiratory syndrome corona virus-2 (SARS-CoV-2). For this, six cows were inoculated with 2 ml of 109.4/ml (TCID50) of the virus. As a control group, three cows were inoculated with the carrier without virus. Specific IgG level against the SARS-CoV-2 was measured before and after immunization in the sera, and in the colostrum and milk following parturition in hyper-immunized cows using indirect Enzyme-linked immunosorbent assay (ELISA). Neutralizing antibodies in the serum and colostrum was measured by a quantitative ELISA. The safety of the product was determined in40 healthy volunteers aged between 18-65 years old (13 females and 27 males) in the phase 1 clinical trial (https://www.irct.ir/trial/51259). No adverse effects were observed in the experimental cows. A very high level of IgG was observed in the first colostrum that sharply decreased in the following 7 days in the milk. The titer of specific neutralizing antibody in the colostrum samples was 69 times higher than the sera. No adverse effects and clinical complications were reported by the authorized ethics committee, and an official certificate on the safety of the product was issued. Beside other strategies, this approach could be used for large-scale and low-cost production of immune components to be used as a nutritional supplement to confront current SARS-CoV-2 and future pandemics. Clinical Trial Registration: [https://www.irct.ir/trial/51259].

Respiratory and GIT tract immune responses of broiler chickens following experimental infection with Newcastle disease's virus.

Newcastle disease causes a lymphoproliferative response in the tracheal and intestinal mucosa of the infected birds. In this study, the Hitchner B1 and I-2 vaccine and challenging of ND field strains were used to evaluate the populations of T lymphocyte subsets infiltrated intestinal and tracheal, also to shed some light on cell-mediated immune response using enzyme-linked immunosorbent assay (ELISA) detecting chicken's serum interferon-γ. Three hundred-day-old broilers were randomly divided into four groups. Groups 1 and 2 received I-2 and B1 vaccines, respectively, while groups 3 and 4 were challenged-unvaccinated and unchallenged-unvaccinated groups. Blood samples were taken from five random chicks and were then tested with ELISA test. Three chicks of each group were euthanized after vaccine administration and also challenging with acute virus. Interferon-γ changes were significant in time (p < 0.001). Totally, there was no significant difference between I-2 and B1 groups. The number of CD3+, CD4+, and CD8+ cells of I-2 and B1 vaccinated group's intestine and the trachea samples was significantly increased compared with the negative control group (p < 0.001). The results indicated the significant increase in CD4+ and CD8+ in intestinal and tracheal tissues, while the level of interferon-γ of the vaccinated group was more than the unvaccinated one. Finding no significant differences between the vaccinated groups indicated the potential of both vaccines in producing CD4+ and CD8+ in the tracheal and intestinal tissues and the equality of interferon-γ production in the sera.

Evaluation of PCR and high-resolution melt curve analysis for differentiation of Salmonella isolates.

Consumption of poultry products contaminated with Salmonella is one of the major causes of foodborne diseases worldwide and therefore detection and differentiation of Salmonella spp. in poultry is important. In this study, oligonucleotide primers were designed from hemD gene and a PCR followed by high-resolution melt (HRM) curve analysis was developed for rapid differentiation of Salmonella isolates. Amplicons of 228 bp were generated from 16 different Salmonella reference strains and from 65 clinical field isolates mainly from poultry farms. HRM curve analysis of the amplicons differentiated Salmonella isolates and analysis of the nucleotide sequence of the amplicons from selected isolates revealed that each melting curve profile was related to a unique DNA sequence. The relationship between reference strains and tested specimens was also evaluated using a mathematical model without visual interpretation of HRM curves. In addition, the potential of the PCR-HRM curve analysis was evaluated for genotyping of additional Salmonella isolates from different avian species. The findings indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Salmonella isolates to determine the serovar/serotype.

Efficacy and transmissibility of Newcastle disease I-2 vaccine strain against a field isolate of virulent ND virus (JF820294.1) in village chicken.

This study was conducted to assess efficacy of heat-stable I-2 vaccine against Newcastle diseases in vaccinated and vaccinated in contact birds group following challenge against virulent Newcastle disease (ND) virus in village chicken. Also, to assess whether birds that have been exposed to vaccine virus-shedding, birds were protected against mortality and clinical signs after infection with a virulent strain of the ND virus (NDV). One hundred fifty one-day-old native chickens were divided into seven groups (4 experimental groups of 30 birds/group and 3 control groups (unvaccinated unchallenged, challenged, and just vaccinated). Birds in experimental groups were vaccinated either via drinking water or as food carrier with thermostable I-2 vaccine and then challenged with virulent isolate of NDV (JF820294.1), and eight birds were added as in-contact birds to vaccinated groups. Following challenge, seven extra birds were added to each group as in contact with vaccinated and challenged birds. Survival rate, clinical signs, necropsy finding, and mean antibody titer were evaluated in different experimental and control groups. Birds vaccinated via drinking water showed 100% survival rate. However, birds vaccinated with food carrier vaccine showed less than 50% survival rate. Based on the results obtained from this study, it can be recommended that I-2 vaccination via drinking water can effectively prevent ND in village chicken, since I-2 strain has been able to transmit to non-vaccinated-sensitive birds more effectively than velogenic NDV.

Vaccination with recombinant 4 × M2e.HSP70c fusion protein as a universal vaccine candidate enhances both humoral and cell-mediated immune responses and decreases viral shedding against experimental challenge of H9N2 influenza in chickens.

As cellular immunity is essential for virus clearance, it is commonly accepted that no adequate cellular immunity is achieved by all available inactivated HA-based influenza vaccines. Thus, an improved influenza vaccine to induce both humoral and cell-mediated immune responses is urgently required to control LPAI H9N2 outbreaks in poultry farms. M2e-based vaccines have been suggested and developed as a new generation of universal vaccine candidate against influenza A infection. Our previous study have shown that a prime-boost administration of recombinant 4×M2e.HSP70c (r4M2e/H70c) fusion protein compared to conventional HA-based influenza vaccines provided full protection against lethal dose of influenza A viruses in mice. In the present study, the immunogenicity and protective efficacy of (r4M2e/H70c) was examined in chickens. The data reported herein show that protection against H9N2 viral challenge was significantly increased in chickens by injection of r4M2e/H70c compared with injection of conventional HA-based influenza vaccine adjuvanted with MF59 or recombinant 4×M2e (r4M2e) without HSP70c. Oropharyngeal and cloacal shedding of the virus was detected in all of the r4M2e/H70c vaccinated birds at 2 days after challenge, but the titer was low and decreased rapidly to reach undetectable levels at 7 days after challenge. Moreover, comparison of protective efficacy against LPAI H9N2 in birds intramuscularly immunized with r4M2e/H70c likely represented the ability of the M2e-based vaccine in providing cross-protection against heterosubtypic H9N2 challenge and also allowed the host immune system to induce HA-homosubtype neutralizing antibody against H9N2 challenge. This protective immunity might be attributed to enhanced cell-mediated immunity, which is interpreted as increased lymphocytes proliferation, increased levels of Th1-type (IFN-γ) and Th2-type (IL-4) cytokines production and increased CD4(+) to CD8(+) ratios, resulting from the injection of four tandem repeats of the ectodomain of the conserved influenza matrix protein M2 (4×M2e) genetically fused to C-terminus of Mycobacterium tuberculosis HSP70 (mHSP70c).

Immunogenicity and protective efficacy of recombinant M2e.Hsp70c (Hsp70(359-610)) fusion protein against influenza virus infection in mice.

New strategies in vaccine development are urgently needed to combat emerging influenza viruses and to reduce the risk of pandemic disease surfacing. Being conserved, the M2e protein, is a potential candidate for universal vaccine development against influenza A viruses. Mycobacterium tuberculosis Hsp70 (mHsp70) is known to cultivate the function of immunogenic antigenpresenting cells, stimulate a strong cytotoxic T lymphocyte (CTL) response, and stop the induction of tolerance. Thus, in this study, a recombinant protein from the extracellular domain of influenza A virus matrix protein 2 (M2e), was fused to the C-terminus of Mycobacterium tuberculosis Hsp70 (Hsp70c), to generate a vaccine candidate. Humoral immune responses, IFN-γ-producing lymphocyte, and strong CTL activity were all induced to confirm the immunogenicity of M2e.Hsp70c (Hsp70(359-610)). And challenge tests showed protection against H1N1 and H9N2 strains in vaccinated groups. Finally these results demonstrates M2e.Hsp70c fusion protein can be a candidate for a universal influenza A vaccine.

Risk factors for detection of bronchial casts, most frequently seen in endemic H9N2 avian influenza infection, in poultry flocks in Iran.

During the last decade, low pathogenic avian influenza (H9N2 subtype) outbreaks with high mortality have been reported in broiler chicken in various countries especially in some Asian and Middle East countries including Iran. Tubular bronchial cast extending to the lower bronchi (BC) is one of the most frequently observed post-mortem lesions in affected broiler chicken during H9N2 low pathogenic avian influenza virus outbreaks. This study was conducted to find out risk factors for high mortality in chickens suffering from respiratory symptoms and showing BC in post-mortem examination. History and general information of the flocks as well as vaccination programs, mortality rate and necropsy findings such as formation of BC in airways were collected for 563 broiler flocks in central part of Fars province, South-West of Iran, during 2001-2006. Results showed that overall mortality rate was 13.52% (95%CI: 12.2-14.8), decreasing from 18% in 2000 to 11% in 2006 (P<0.01). Corresponding measures for BC were 22% (95%CI: 18-26) with no significant decreasing trend during the study period (P=0.14). Multivariable logistic regression analysis showed that infectious bronchitis (IB) vaccination, flock size and geographical location were significantly associated with the observation of BC (P<0.05). Odds of BC observation in flocks with history of IB vaccination was seven times higher than flocks without vaccination, and in small flocks was nearly half of the large flocks. In conclusion, IB vaccine can be one of the candidate risk factors for enhancing the virulence of low pathogenic H9N2 virus in the fields.

Prokaryotic expression and characterization of avian influenza A virus M2 gene as a candidate for universal recombinant vaccine against influenza A subtypes; specially H5N1 and H9N2.

The conserved M2 protein of influenza A virus is considered as a promising candidate target for a broad-spectrum, recombinant influenza A vaccine. In the present study, the open reading frame (ORF) of avian influenza A/chicken/Iran/101/1998 (H9N2) M2 gene was amplified then cloned in pAED4, prokaryotic expression vector. M2 protein was produced through the expression of this recombinant expression vector (pAED4-M2) in E. coli BL21 (DE3) strain. The expressed M2 protein was analyzed on SDS-PAGE. Western blot assay was used to examine the immunoreaction of the expressed protein using commercial polyclonal anti-M2 antibody. The antigenicity and biological activity of the recombinant protein was also qualitatively detected on infected MDCK cells surface by immunofluorescence assay using rabbit's immunized antiserum. So, according to the sequence alignment based on the mentioned isolate and the result of immunoassay reaction, it seems recombinant vaccine based on A/chicken/Iran/101/1998(H9N2) M2 protein isolate might cover majority of influenza A virus strains specially H5 and H9 circulating in Iran and neighbor regions significantly.

Pathogenesis of highly pathogenic avian influenza A/turkey/Turkey/1/2005 H5N1 in Pekin ducks (Anas platyrhynchos) infected experimentally.

Asian H5N1 (hereafter referred to as panzootic H5N1) highly pathogenic avian influenza (HPAI) virus has caused large numbers of deaths in both poultry and wild-bird populations. Recent isolates of this virus have been reported to cause disease and death in commercial ducks, which has not been seen with other HPAI viruses. However, little is known about either the dissemination of this H5N1 within the organs or the cause of death in infected ducks. Nineteen 4-week-old Pekin ducks were infected with 10(6.7) median egg infectious doses of HPAI A/turkey/Turkey/1/05 (H5N1, clade 2.2) in 0.1ml via the intranasal and intraocular routes. Cloacal and oropharyngeal swabs were taken daily before three animals were selected randomly and killed humanely for postmortem examination, when samples of tissues were taken for real-time reverse transcriptase-polymerase chain reaction, histopathological examination and immunohistochemistry. Clinical signs were first observed 4 days post infection (d.p.i.) and included depression, reluctance to feed, in-coordination and torticollis resulting in the death of all the birds remaining on 5d.p.i. Higher levels of virus shedding were detected from oropharyngeal swabs than from cloacal swabs. Real-time reverse transcriptase-polymerase chain reaction and immunohistochemistry identified peak levels of virus at 2d.p.i. in several organs. In the spleen, lung, kidney, caecal tonsils, breast muscle and thigh muscle the levels were greatly reduced at 3d.p.i. However, the highest viral loads were detected in the heart and brain from 3d.p.i. and coincided with the appearance of clinical signs and death. Our experimental results demonstrate the systemic spread of this HPAI H5N1 virus in Pekin ducks, and the localization of virus in the brain and heart tissue preceding death.

Natural cases and an experimental study of H9N2 avian influenza in commercial broiler chickens of Iran.

Since 1998, an epidemic of avian influenza has occurred in the Iranian poultry industry. The agent was pathotyped as non-highly pathogenic and subtyped as an H9N2 avian influenza virus. Therefore it did not require eradication. However, frequent incidences of high mortality were observed commonly on broiler farms. No other species of bird were affected. The circulation of the virus and mixed infection with other respiratory pathogens, particularly infectious bronchitis virus and Mycoplasma gallisepticum, were incriminated in the high mortality on poultry farms and resulting great economic losses. Clinical signs in both field and experimental studies included swelling of the periorbital tissues and sinuses, nasal and ocular discharge, and severe respiratory distress. However, in the experimental study, the mortality rate was much lower than in the natural outbreak. Gross lesions identified included extensive congestion of the respiratory tissues, and exudation with cast formation in the tracheal bifurcation, which extended to the secondary bronchi. Severe necrotizing tracheatis was the predominate histological lesion. Ultrastructurally, orthomyxovirus-like particles were identified in the inoculum used for the experimental study. An inactivated H9N2 avian influenza vaccine prevented mortality in experimentally challenged chickens.