Comparative Molecular Virulence Typing and Antibiotic Resistance of Campylobacter Species at the Human-Animal-Environment Interface.

This study holds significant importance due to its focus on Campylobacter, the leading bacterial cause of gastroenteritis worldwide, responsible for ∼96 million cases each year. By investigating the prevalence of both Campylobacter jejuni and Campylobacter coli in humans, animals, and the environment, this research sheds light on the broader impact of these pathogens, which can harm both human and animal populations. Traditional microbiological methods were implemented followed by optimized multiplex polymerase chain reaction targeting 16S rDNA and virulence gene markers by using specific primers. The findings revealed that a total of 219 Campylobacter isolates were recovered from 528 collected specimens from human, animal, and environmental sources. Campylobacter species showed a prevalence of 41.5%, with C. jejuni accounting for 53% and C. coli for 47%. Antimicrobial resistance rates were high, with tetracycline at 89%, ceftriaxone at 75%, cefotaxime at 70%, erythromycin at 69%, nalidixic acid at 54%, ciprofloxacin at 39%, and gentamicin at 23%. Commonly prevalent virulence-associated genes observed in the Campylobacter were cadF at 93%, flaA at 91%, cdtB at 88%, cheY at 86%, sodB at 78%, and iamA at 32%. The study confirmed multidrug-resistant Campylobacter prevalence at the human-animal-environment interface, harboring virulence-associated genes with potential harm to humans. Data analysis showed a nonsignificant (p ≥ 0.05) correlation between virulence genes and antibiotic susceptibility. To effectively manage Campylobacter infections, a multifaceted strategy incorporating preventative interventions at different levels is required. This strategy should take into account practicability, effectiveness, and sustainability while strengthening surveillance systems and addressing the economics of disease prevention.

Development of egg yolk-based polyclonal antibodies and immunoprophylactic potential of antigen-antibody complex against infectious bursal disease.

A study conducted in the Faisalabad district sampled 50 cases across five IBD outbreaks, revealing an alarming 80 % infection rate among poultry. Our research focused on developing an immune complex (Antigen-antibody complex) with potential immunoprophylactic benefits to counter this formidable threat. Our study was based on producing egg yolk-derived polyclonal antibodies (IgY) targeting IBDV. Commercial layer birds were immunized with inactivated IBDV, yielding IgY antibodies extracted from their eggs exhibited substantially higher and more enduring antibody titers, with a geometric mean titer of 104. Further research involved the creation of an immune complex (ICx) where antigen was extracted from infected bursae tissues. The immunogenic response of ICx was assessed in poultry birds after a challenge with a virulent strain of IBD virus and compared to a conventional IBDV vaccine in poultry. Results revealed significantly higher and more enduring antibody titers induced by the ICx, offering enhanced protective immunity against the IBDV challenge, as evidenced by lower Bursa to bodyweight ratios in vaccinated birds.

Molecular identification and cross-immunogenic study on two field isolates of Mycoplasma synoviae isolated from broilers in five districts of Khyber Pakhtunkhwa.

Background: Mycoplasma synoviae (MS) is an important poultry pathogen causing heavy economic losses Worldwide. Subclinical persistence of this pathogen is the major issue to control its prevalence. Aim: This study aimed to determine the molecular and cross-immunogenicity of MS among broilers in five Districts of Khyber Pakhtunkhwa (KP). Methods: This study was conducted by collecting 434 specimen samples from 40 broiler farms and desi poultry in five districts of KP. Specimen samples from the broiler birds (n = 150), broiler farm environment (n = 264), and desi poultry birds (n = 20) were aseptically collected and serially passaged in Modified Frey's broth. The homologous and heterologous antibody reactions were studied in rabbits. Before inoculation into rabbits, the MS isolates were inactivated by formalin and adjuvanted with Montanide. Results: The overall turbidity prevalence in Frey's broth was observed as 109/434 (25.11%) samples, and these turbidity-positive samples were shifted on Frey's agar. After the appearance of classic fried egg colonies, the Biochemical confirmation was supported by the production of catalase and phosphatase, reduction of tetrazolium, film and spot assay, and fermentation of glucose for species differentiation in avian mycoplasma. The MS prevalence percentage was recorded as 2% (9/434) through biochemical tests. The PCR results showed 0.5% MS prevalence with two field isolates (named MS-1 and MS-2). Both MS-1 and MS-2 field isolates showed similar values (42.2) of homologous geometric mean titer (GMT). While the heterologous GMT for MS-1 serum against MS-2 isolate was lower (27.9) as compared to MS-2 serum against MS1 isolate (38.9). No titer was detected in the control group (Group-III). Conclusion: In conclusion, the results indicated the existence of MS in broiler birds and high homologous titers recorded between field isolates, which is a perpetual menace to poultry.

Development of a Lateral-Flow Immunochromatographic Strip for the Detection of Oxytetracycline Residues in Biological Fluids.

Oxytetracycline (OTC) is extensively used in veterinary medicine and for growth promotion around the globe. The indiscriminate use of OTC in food-producing animals leaves residues in animal products. The presence of these residues in animal products causes economic losses and harmful effects on consumers. Different regulatory bodies set maximum residue limits (MRLs) for different tetracyclines. To avoid harmful effects, there is a need for a simple, fast, and economical method for the screening of animal products. In this study, a fast, economical, and user-friendly lateral-flow immunochromatographic (LFIC) assay based on gold nanoparticles (AuNPs) was developed to detect the presence of OTC residues in biological fluids. AuNPs provided visual results as red lines in 6-15 min. Polyclonal rabbit IgG antibodies were produced using the immunogen of OTC. These antibodies were purified by the combined ammonium sulfate-octanoic acid precipitation method. Antibodies were conjugated to AuNPs as recognition biomolecules. A LFIC strip was optimized using borate buffer spiked with different concentrations of the OTC. The visual limit of detection (LOD) in different biological samples (milk, serum, and urine) was determined using samples spiked with OTC. The LOD was found to be 15 µg/L, which is very low from the MRL (100 µg/L) set by different regulatory authorities. This LFIC strip can be used to detect OTC residues in biological fluids for point-of-care testing (POCT). These strips are easy to use, cost-effective, and portable and provide quick results without the use of laboratory instruments.

Association between antimicrobial consumption and resistance rate of Escherichia coli in hospital settings.

AIMS: This study aims to quantify antimicrobial consumption (AMC) and antimicrobial resistance (AMR) in tertiary care hospitals in Pakistan. METHODS AND RESULTS: This observational study was conducted at two tertiary care hospitals of Pakistan over a 1-year period. The AMC and resistance patterns of Escherichia coli isolates collected from hospitals were observed and analysed using the pearson correlation coefficient. AMC in hospitals varied between 0.00186 and 0.72 (Mean = 0.12 ± 0.191) Defined Daily Dose (DDDs)/1000 patient-days. Fluoroquinolones were the most consumed antimicrobial followed by penicillins, cephalosporins, carbapenems, and aminoglycosides. Antimicrobial-resistant rates in hospitals varied between 92.2% and 34.6%. The highest resistance rate was observed for moxifloxacin, followed by ampicillin, cefotaxime, amoxicillin, ceftriaxone, ciprofloxacin, amoxiclav, and amikacin. Statistically significant association was found between AMC and resistance rate for ampicillin (r = 0.78, P = 0.032), cefotaxime (r = 0.87, P = 0.012), ceftriaxone (r = 0.67, P = 0.042), and ciprofloxacin (r = 0.63, P = 0.031). Additionally, there was a significant association between fluoroquinolone consumption and the resistance rate of third generation cephalosporins (r = 0.61, P = 0.032), and significance was also found when all antimicrobials were combined into 1 analysis (r = 0.721, P = 0.032). CONCLUSION: This data documented a significant association between AMC and resistant rates for multiple antimicrobial agents.

Potential Mechanisms of Transmission of Tick-Borne Viruses at the Virus-Tick Interface.

Ticks (Acari; Ixodidae) are the second most important vector for transmission of pathogens to humans, livestock, and wildlife. Ticks as vectors for viruses have been reported many times over the last 100 years. Tick-borne viruses (TBVs) belong to two orders (Bunyavirales and Mononegavirales) containing nine families (Bunyaviridae, Rhabdoviridae, Asfarviridae, Orthomyxovirida, Reoviridae, Flaviviridae, Phenuviridae, Nyamiviridae, and Nairoviridae). Among these TBVs, some are very pathogenic, causing huge mortality, and hence, deserve to be covered under the umbrella of one health. About 38 viral species are being transmitted by <10% of the tick species of the families Ixodidae and Argasidae. All TBVs are RNA viruses except for the African swine fever virus from the family Asfarviridae. Tick-borne viral diseases have also been classified as an emerging threat to public health and animals, especially in resource-poor communities of the developing world. Tick-host interaction plays an important role in the successful transmission of pathogens. The ticks' salivary glands are the main cellular machinery involved in the uptake, settlement, and multiplication of viruses, which are required for successful transmission into the final host. Furthermore, tick saliva also participates as an augmenting tool during the physiological process of transmission. Tick saliva is an important key element in the successful transmission of pathogens and contains different antimicrobial proteins, e.g., defensin, serine, proteases, and cement protein, which are key players in tick-virus interaction. While tick-virus interaction is a crucial factor in the propagation of tick-borne viral diseases, other factors (physiological, immunological, and gut flora) are also involved. Some immunological factors, e.g., toll-like receptors, scavenger receptors, Janus-kinase (JAK-STAT) pathway, and immunodeficiency (IMD) pathway are involved in tick-virus interaction by helping in virus assembly and acting to increase transmission. Ticks also harbor some endogenous viruses as internal microbial faunas, which also play a significant role in tick-virus interaction. Studies focusing on tick saliva and its role in pathogen transmission, tick feeding, and control of ticks using functional genomics all point toward solutions to this emerging threat. Information regarding tick-virus interaction is somewhat lacking; however, this information is necessary for a complete understanding of transmission TBVs and their persistence in nature. This review encompasses insight into the ecology and vectorial capacity of tick vectors, as well as our current understanding of the predisposing, enabling, precipitating, and reinforcing factors that influence TBV epidemics. The review explores the cellular, biochemical, and immunological tools which ensure and augment successful evading of the ticks' defense systems and transmission of the viruses to the final hosts at the virus-vector interface. The role of functional genomics, proteomics, and metabolomics in profiling tick-virus interaction is also discussed. This review is an initial attempt to comprehensively elaborate on the epidemiological determinants of TBVs with a focus on intra-vector physiological processes involved in the successful execution of the docking, uptake, settlement, replication, and transmission processes of arboviruses. This adds valuable data to the existing bank of knowledge for global stakeholders, policymakers, and the scientific community working to devise appropriate strategies to control ticks and TBVs.

Idiotype/anti-idiotype antibodies: as a glorious savior in COVID-19 pandemics.

The idiotype network is experimentally modified to provide protective immunity against various microbial pathogens. Both internal and non-internal image-idiotype antibodies can trigger specific immune responses to antigens. The current outbreak of Severe Acute Respiratory Syndrome 2 (SARS-2) has provided a great opportunity to take advantage of idiotype / anti-idiotype antibodies as a protective regimen when no approved vaccine is available on earth. The current review identifies successful applications of idiotype/ anti-idiotype antibodies in various viral diseases and highlights their importance in COVID-19 pandemics. In the absence of vaccines and targeted therapies, polyclonal idiotype/ anti-idiotype antibodies against the viral structure may be a potential approach to the prevention and treatment of COVID-19 patients.

Molecular Characterization and Demographic Study on Infectious Bursal Disease Virus in Faisalabad District.

The re-emergence of virulent strains of the Infectious Bursal Disease Virus (IBDV) leads to significant economic losses of poultry industry in Pakistan during last few years. This disease causes the infection of bursa, which leads to major immune losses. A total number of 30 samples from five IBD outbreaks during the period of 2019-20 were collected from different areas of Faisalabad district, Pakistan and assayed by targeting the IBD virus VP2 region through RT-PCR. Among all the outbreaks, almost 80% of poultry birds were found positive for the IBDV. The bursa tissues were collected from the infected birds and histopathological examination of samples revealed severe lymphocytic depletion, infiltration of inflammatory cells, and necrosis of the bursa of Fabricius (BF). Positive samples were subjected to re-isolation and molecular characterization of IBDV. The Pakistan IBDV genes were subjected to DNA sequencing to determine the virus nucleotide sequences. The sequences of 100 Serotype-I IBDVs showing nearest homology were compared and identified with the study sequence. The construction of the phylogenetic tree for nucleotide sequences was accomplished by the neighbor-joining method in MEGA-6 with reference strains. The VP2 segment reassortment of IBDVs carrying segment A were identified as one important type of circulating strains in Pakistan. The findings indicated the molecular features of the Pakistan IBDV strains playing a role in the evolution of new strains of the virus, which will contribute to the vaccine selection and effective prevention of the disease.

Type I IFNs: A Blessing in Disguise or Partner in Crime in MERS-CoV-, SARS-CoV-, and SARS-CoV-2-Induced Pathology and Potential Use of Type I IFNs in Synergism with IFN-γ as a Novel Antiviral Approach Against COVID-19.

Since the end of 2019, the emergence of novel coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has accelerated the research on host immune responses toward the coronaviruses. When there is no approved drug or vaccine to use against these culprits, host immunity is the major strategy to fight such infections. Type I interferons are an integral part of the host innate immune system and define one of the first lines of innate immune defense against viral infections. The in vitro antiviral role of type I IFNs against Middle East respiratory syndrome coronavirus (MERS-CoV) and SARS-CoV (severe acute respiratory syndrome coronavirus) is well established. Moreover, the involvement of type I IFNs in disease pathology has also been reported. In this study, we have reviewed the protective and the immunopathogenic role of type I IFNs in the pathogenesis of MERS-CoV, SARS-CoV, and SARS-CoV-2. This review will also enlighten the potential implications of type I IFNs for the treatment of COVID-19 when used in combination with IFN-γ.

Isolation and identification of Saccharomyces cerevisiae for fermentation of rice polishing in Livestock feeding.

Biomass of Saccharomyces cerevisiae wasenhancedin rice polishing by fermentation to increase protein contents of feedfor its use in livestock. Broth culture of Saccharomyces cerevisiae (2.6×108CFU/mL) was prepared from culture obtained by continuous streaking. The isolated culture was identified morphologically by Gram staining and confirmed by biochemical characteristics. Rice polishing was sieved to remove larger particles. Then it was distributed to 4 treatments in triplicates. Treatments were represented as rice polishing (RP), rice polishing plus Saccharomyces culture (RPSC), rice polishing plus ammonium sulphate (RPAS), rice polishing plus Saccharomyces culture plus ammonium sulphate (RPSCAS).Fermentation was provided for 144 hours at 320C,while samples were collected after every 24 hours. Samples were dried, ground and subjected to proximate analysis. It was observed that protein content was increased from 11% to 21.51% and maximum increment was obtained after 144 hours of incubation in RPSC treatment. Ether extract and ash were increased from 14% and 10% to 16.96% and 11.11% in RPSCAS respectively. A significant reduction in neutral detergent fiber was observed after fermentation. It is concluded that Saccharomyces cerevisiae has potential to improve mineral and protein contents of rice polishing by fermentation process with or without addition of nitrogen source.

Comparative Study of Protection against Newcastle Disease in Young Broilers Administered Natural Chicken Alpha Interferon via Oral and Intramuscular Routes.

Despite extensive vaccination approaches, Newcastle disease (ND) remains a permanent threat to the poultry industry worldwide. Besides vaccination, there is a burgeoning demand for new antivirals for use in interventions to control ND. One strategy is to strengthen the host innate immunity via host-derived innate immune proteins. Type I interferons define one of the first lines of innate immune defense against viral infections. Chicken interferon alpha (chIFN-α) is one of the potent cytokines that trigger antiviral responses. In the current study, we investigated the therapeutic effect of natural chIFN-α administered via oral and intramuscular (i.m.) routes against ND in broiler chickens. Our results showed that the level of protection against ND in response to chIFN-α therapy was dependent on the route and dose of IFN administration. A better therapeutic effect was observed in chickens treated with chIFN-α via the oral route than in those treated via the i.m. route. Regardless of the administration route, double-dose chIFN-α (2,000-U) treatments provided better protection than single-dose (1,000-U) treatments. However, complete protection against ND was achieved in birds treated with repeated doses of chIFN-α via the oral route. Histopathology of trachea, proventriculus, spleen, and liver showed a significant improvement in ND-induced degenerative changes in double-dose IFN-treatment groups compared to single-dose groups. Results of the hemagglutination test demonstrated a decrease in ND virus (NDV) titer in IFN-treated groups. Also, double doses of chIFN-α via oral route resulted in early recovery in weight gain. We propose that chIFN-α therapy via oral route could be an important therapeutic tool to control NDV infection in chicken.IMPORTANCE Newcastle disease (ND) is an economically important contagious disease of wild and domestic birds worldwide. The disease causes severe economic losses in terms of production due to high mortality and morbidity in nonvaccinated chickens. Despite extensive vaccination approaches, Newcastle disease (ND) remains a permanent threat to the poultry industry worldwide. In the current study, we used natural chicken IFN-α as an innate immune modulator to counteract ND in chickens. We report that chIFN-α is effective in protecting the chickens against ND and also prevents shedding of the virus, which can then prevent further spread of the disease. We propose that in addition to vaccination, chIFN-α therapy could be an effective option for controlling ND in areas of endemicity.

Anti-chicken type I IFN countermeasures by major avian RNA viruses.

Chicken type I interferons (type I IFNs) are key antiviral players of the chicken innate immune system and are considered potent antiviral agents against avian viral pathogens. Chicken type I IFNs are divided into three subtypes namely, chIFN-α, chIFN-ß, and chIFN-κ. Viral pathogen-associated molecular patterns (PAMPs) recognized by their corresponding specific PRRs (pattern recognition receptors) induce the expression of chicken type I IFNs. Interaction of chicken type I IFNs with their subsequent IFN receptors results in the activation of the JAK-STAT pathway, which in turn activates hundreds of chicken interferon-stimulated genes (chISGs). These chISGs establish an antiviral state in neighboring cells and prevent the replication and dissemination of viruses within chicken cells. Chicken type I IFNs activate different pathways that constitute major antiviral innate defense mechanisms in chickens. However, evolutionary mechanisms in viruses have made them resistant to these antiviral players by manipulating host innate immune pathways. This review focuses on the underlying molecular mechanisms employed by avian RNA viruses to counteract chicken type I IFNs and chISGs through different viral proteins. This may help to understand host-pathogen interactions and the development of novel therapeutic strategies to control viral infections in poultry.

Novel Coronavirus disease 2019 (COVID-19): new challenges and new responsibilities in developing countries.

Novel coronavirus disease 2019 (COVID-19) is caused by the SARS-CoV-2 virus, which belongs to the genus Coronaviridae with its high mutation rate. From the current perspective, we discuss the current status of COVID-19, new challenges, and potential interventions to control the pandemic in developing counties such as Pakistan.

Enhancement in humoral response against inactivated Newcastle disease vaccine in broiler chickens administered orally with plant-derived soyasaponin.

The present study aimed to evaluate the immunopotentiating effect of plant-derived soyasaponin and its immunogenicity in chickens challenged with Newcastle disease virus (NDV). Soyasaponin was extracted from soybean seeds and detected using the phytochemical tests, followed by quantification through the dry-weight method. One-day-old broiler chicks (n = 90) were divided into 3 groups, named as A, B, and C. Group A birds were orally administrated with soyasaponin (5 mg/kg), followed by immunization with inactivated ND vaccine intramuscularly (IM), whereas group B birds were vaccinated with inactivated ND vaccine alone. Group C birds were kept unvaccinated. A booster dose on day 21 was also administered IM to group A and B birds. At day 35, all 3 groups were challenged with NDV. To determine the immunogenicity potential of soyasaponin, antibody titer was measured using the hemagglutination inhibition test before and after the NDV challenge. Histochemical examination was performed to determine the pathological changes associated with NDV infection. Foam formation and hemolytic activity confirmed the presence of saponin in soya bean extract. Group A birds showed a higher antibody response compared with group B and C birds. The disease challenge study showed that soyasaponin-adjuvanted NDV vaccine provided complete protection to group A birds against ND. Moreover, no side effects of soyasaponin were observed on the growth performance of birds during the experiment. Therefore, we can conclude that soyasaponin is a potential immunogenic agent and therefore could be a promising candidate to launch a protective humoral response against ND in chickens.

Web of interferon stimulated antiviral factors to control the influenza A viruses replication.

Influenza viruses cause mild to severe infections in animals and humans worldwide with significant morbidity and mortality. Infection of eukaryotic cells with influenza A viruses triggers the induction of innate immune system through the interaction between pattern recognition receptors (PRRs) and pathogen associated molecular patterns (PAMPs), which culminate in the induction of interferons (IFNs). Consequently, IFNs bind to their cognate receptors on the cellular membrane and activate the signaling pathway for transcriptional regulation of interferon-stimulated genes (ISGs) through Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway. Cumulative actions of these ISGs establish an antiviral state of the host. Several ISGs have been described, which play critical roles to inhibit the infection and replication of influenza A viruses at multiple steps of virus life cycle. In this review, the dynamics and redundancy of these ISGs against influenza A viruses are discussed. Additionally, current understanding and molecular mechanisms that are underlying the roles of ISGs in pathogenesis of influenza virus are critically reviewed.

Comprehensive network map of transcriptional activation of chicken type I IFNs and IFN-stimulated genes.

Chicken type I interferons (type I IFNs) are key antiviral players of the chicken immune system and mediate the first line of defense against viral pathogens infecting the avian species. Recognition of viral pathogens by specific pattern recognition receptors (PRRs) induce chicken type I IFNs expression followed by their subsequent interaction to IFN receptors and induction of a variety of IFN stimulated antiviral proteins. These antiviral effectors establish the antiviral state in neighboring cells and thus protect the host from infection. Three subtypes of chicken type I IFNs; chIFN-α, chIFN-ß, and a recently discovered chIFN-κ have been identified and characterized in chicken. Chicken type I IFNs are activated by various host cell pathways and constitute a major antiviral innate defense in chicken. This review will help to understand the chicken type 1 IFNs, host cellular pathways that are involved in activation of chicken type I IFNs and IFN stimulated antiviral effectors along with the gaps in knowledge which will be important for future investigation. These findings will help us to comprehend the role of chicken type I IFNs and to develop different strategies for controlling viral infection in poultry.

Evaluation of a Salmonella Strain Isolated from Honeybee Gut as a Potential Live Oral Vaccine Against Lethal Infection of Salmonella Typhimurium.

In this research, Salmonella species were isolated from the animal, insect and human enteric sources in Faisalabad, Punjab, Pakistan. These species were characterized by different microbiological and molecular techniques including polymerase chain reaction (PCR) by amplification of the 16S rRNA gene. Furthermore, sequencing of the amplicons confirmed all ten isolates as Salmonella strains. The antigenic cross-reactivity was found maximum between the HB1 (strain isolated from honeybee) antiserum and its antigen with an antibody titer of 1:128, while the HB1 antiserum showed a cross-reactive titer range of 1:8 to 1:64. On the basis of the highest geometric mean titer (GMT) shown by the antiserum of the HB1 antigen, it was selected as the best candidate for a cross-reactive live Salmonella oral antigen. Moreover, the HB1 antigen was used a live oral antigen (1 × 1010 CFU/ml) in a safety test in rabbits and proved to be avirulent. During the animal trial, three different oral doses of the HB1 live oral antigen were evaluated in four different rabbits' groups (R1, R2, R3, and R4). The dose number 2 of 0.5 ml (two drops orally and repeated after one week) gave the best GMT measured by indirect hemagglutination (IHA) as compared to the other two doses, while R4 group was kept as control. Results of the challenge protection test also validated the efficacy of the double dose of the HB1 live vaccine, which gave the highest survival percentage. Results of this study lay the foundation for a potential cross-reactive live oral Salmonella vaccine that has proved to be immunogenic in rabbits.In this research, Salmonella species were isolated from the animal, insect and human enteric sources in Faisalabad, Punjab, Pakistan. These species were characterized by different microbiological and molecular techniques including polymerase chain reaction (PCR) by amplification of the 16S rRNA gene. Furthermore, sequencing of the amplicons confirmed all ten isolates as Salmonella strains. The antigenic cross-reactivity was found maximum between the HB1 (strain isolated from honeybee) antiserum and its antigen with an antibody titer of 1:128, while the HB1 antiserum showed a cross-reactive titer range of 1:8 to 1:64. On the basis of the highest geometric mean titer (GMT) shown by the antiserum of the HB1 antigen, it was selected as the best candidate for a cross-reactive live Salmonella oral antigen. Moreover, the HB1 antigen was used a live oral antigen (1 × 1010 CFU/ml) in a safety test in rabbits and proved to be avirulent. During the animal trial, three different oral doses of the HB1 live oral antigen were evaluated in four different rabbits' groups (R1, R2, R3, and R4). The dose number 2 of 0.5 ml (two drops orally and repeated after one week) gave the best GMT measured by indirect hemagglutination (IHA) as compared to the other two doses, while R4 group was kept as control. Results of the challenge protection test also validated the efficacy of the double dose of the HB1 live vaccine, which gave the highest survival percentage. Results of this study lay the foundation for a potential cross-reactive live oral Salmonella vaccine that has proved to be immunogenic in rabbits.

Immune Modulatory Potential of Anti-idiotype Antibodies as a Surrogate of Foot-and-Mouth Disease Virus Antigen.

The immunoprophylactic potential of anti-idiotype (anti-id) foot-and-mouth disease (FMD) antigen (Ag) was evaluated in the calves. The idiotype antibodies (Ab1) were produced in experimental goats by injecting inactivated FMD virus. The Fab (fragment antigen binding) of Ab1 was injected into the layer birds to raise anti-id antibodies (Ab2). The Ab2 was purified from egg yolks. The Fab component of Ab2 was emulsified in Montanide (1:1) and used as a surrogate of FMD virus. The immune response to Montanide adjuvanted monovalent and trivalent anti-id FMD virus antigen was determined in mice. The comparative immune potentiation potentials of Montanide adjuvanted trivalent anti-id FMD virus antigen and trivalent FMD vaccine were determined in mice and calves. Montanide adjuvanted monovalent anti-id FMD virus antigens produced mean Ab titers of 78.80%, 81.30%, and 81.20% for serotypes A, Asia 1, and O, respectively, at 45 days postimmunization (p.i.) in mice. Montanide adjuvanted trivalent anti-id FMD Ag in mice produced the highest Ab titer, 81.60%, at day 45 compared to the 77.50% titer measured for Montanide adjuvanted FMD vaccine at day 45 p.i. A slow decrease of 1% to 2% was recorded for the Ab titer of Montanide adjuvanted trivalent anti-id FMD virus antigen in mice at day 60. In calves, the titer corresponding to the immune response seen with Montanide adjuvanted trivalent anti-id FMD virus antigen (80%) was persistent whereas the titer of Montanide adjuvanted FMD vaccine decreased to 74% at day 60 p.i. Anti-id FMD virus antigen induced a strong and persistent immunogenic response in terms of Ab titer compared to the inactivated virus vaccine. Anti-id FMD virus antigen may serve as a surrogate of FMD virus vaccine.IMPORTANCE Foot-and-mouth disease (FMD) is a contagious viral disease of animals. Multiple serotypes and antigenic variation in the viral genome are probably the factors that reduce control of the disease. Currently, the vaccines employed against FMD use killed virus. The inactivation or killing of the virus makes it less immunogenic and reduces its immunoprophylactic potential. To cope with this situation, the present study was designed, anti-idiotype FMD virus antigen was prepared, and the immunogenic potential of the antigen was compared to that of commercial killed-virus vaccines. The overall results showed that a persistent and strong immune response occurred with anti-idiotype FMD virus antigen. Thus, anti-idiotype FMD virus antigen may serve as a potential surrogate of FMD virus vaccines.

Assessing the capability of Fourier transform infrared spectroscopy in tandem with chemometric analysis for predicting poultry meat spoilage.

BACKGROUND: Use of traditional methods for determining meat spoilage is quite laborious and time consuming. Therefore, alternative approaches are needed that can predict the spoilage of meat in a rapid, non-invasive and more elaborative way. In this regard, the spectroscopic techniques have shown their potential for predicting the microbial spoilage of meat-based products. Consequently, the present work was aimed to demonstrate the competence of Fourier transform infrared spectroscopy (FTIR) to detect spoilage in chicken fillets stored under aerobic refrigerated conditions. METHODS: This study was conducted under controlled randomized design (CRD). Chicken samples were stored for 8 days at 4 + 0.5 °C and FTIR spectra were collected at regular intervals (after every 2 days) directly from the sample surface using attenuated total reflectance during the study period. Additionally, total plate count (TPC), Entetobacteriaceae count, pH, CTn (Color transmittance number) color analysis, TVBN (total volatile basic nitrogen) contents, and shear force values were also measured through traditional approaches. FTIR spectral data were interpreted through principal component analysis (PCA) and partial least square (PLS) regression and compared with results of traditional methods for precise estimation of spoilage. RESULTS: Results of TPC (3.04-8.20 CFU/cm2), Entetobacteriaceae counts (2.39-6.33 CFU/cm2), pH (4.65-7.05), color (57.00-142.00 CTn), TVBN values (6.72-33.60 mg/100 g) and shear force values (8.99-39.23) were measured through traditional methods and compared with FTIR spectral data. Analysis of variance (ANOVA) was applied on data obtained through microbial and quality analyses and results revealed significant changes (P < 0.05) in the values of microbial load and quality parameters of chicken fillets during the storage. FTIR spectra were collected and PCA was applied to illuminate the wavenumbers potentially correlated to the spoilage of meat. PLS regression analysis permitted the estimates of microbial spoilage and quality parameters from the spectra with a fit of R2 = 0.66 for TPC, R2 = 0.52 for Entetobacteriaceae numbers and R2 = 0.56 for TVBN analysis of stored broiler meat. DISCUSSION: PLS regression was applied for quantitative interpretation of spectra, which allowed estimates of microbial loads on chicken surfaces during the storage period. The results suggest that FTIR spectra retain information regarding the spoilage of poultry meat. CONCLUSION: The present work concluded that FTIR spectroscopy coupled with multivariate analysis can be successfully used for quantitative determination of poultry meat spoilage.

Removal of copper from an electroplating industrial effluent using the native and modified spirogyra.

In the present study, biosorption behavior of a green filamentous alga, spirogyra in its native and modified states was investigated for copper removal from an electroplating industrial effluent. For this, the effluent containing 194 mg·L-1 Cu2+ in sulfate medium was contacted with both forms of spirogyra, under the parametric variations of effluent pH, adsorbent dosage, contact time, and sorption temperature. The study revealed spirogyra as a prominent candidate for removing contaminant metal cation; however, at the same condition, biosorption capacity of modified biomass in gel form was higher than the native spirogyra. At the optimized condition with 6 g sorbent dosage treated to 100 mL effluent for 30 min at pH 6.0 and temperature 20 °C, the maximum 82.8% and 96.4% copper could be adsorbed by the native and modified spirogyra, respectively. The batch sorption data using native biomass followed pseudo-first-order kinetic; exhibiting the multilayer sorption mechanism via surface diffusion could be defined by the Freundlich model. In contrast, the sulfuric acid treated modified spirogyra followed pseudo-second-order kinetics and intra particle diffusion as the rate-limiting step.