Plant Extracts: Potential Alternative Treatment for Bovine Mastitis Causing Pathogen Staphylococcus aureus

Bovine mastitis is a significant disease affecting dairy herds worldwide. Mastitis can be characterized by physical, chemical, and bacteriological changes in milk and various pathological changes in the glandular tissues. This disease can invariably affect the health status of cattle and eventually have a direct economic impact on the dairy industry. Mastitis can be caused by the interaction of pathogens and their environment, and one of the disease-causing pathogens, Staphylococcus aureus, remains the leading cause of mastitis. Treatment is directed towards the use of broad-spectrum antimicrobials. However, with the threat of antibiotic-resistant pathogens, alternative treatments are being explored. The use of plants with ethnoveterinary origins can be promising in the search for novel therapeutic regimens. This review focuses on various studies using plant extracts as a possible alternative treatment for this specific bovine-causing pathogen, Staphylococcus aureus. Several studies that were conducted will serve as preliminary data in the development of alternative treatments for bovine mastitis

SCREENING OF THE ACID MEAT CONDITION IN THE RENDEMENT NAPOLE GENE USING POLYMERASE CHAIN REACTION - RESTRICTION FRAGMENT LENGTH POLYMORPHISM

A mutation in the rendement napole (RN) gene causes the acid meat condition which results to poor meat quality due to its reduced water holding capacity, low pH, pale color, reduced processing and cooking yield due to increased drip, and strong metallic taste. This study was conducted to detect the mutation in the RN gene in 535 commercial breeder pigs from the Philippines. Blood collection was done then subjected to DNA extraction and genotyping using polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) using the enzyme BsrBI, then validated by DNA sequencing. Results revealed that 97.01% of the breeder pigs did not have the mutation in their RN gene, while 2.69% had at least one copy of the defective allele in their gene. The acid meat condition has only been previously detected in the Hampshire breed whereas this study found the mutations predominantly in Pietrain and Landrace breed they were classified as normal (rn/rn), heterozygous mutants (RN/rn), and homozygous mutants (RN/RN) which allowed breeding systems to be developed ensuring that all offspring are free of the defect.  This genetic screening will help in detecting the presence of the defect in a given swine population and reduce the unwanted effects on meat quality thus increasing its market value

Emerging Infectious Diseases in Water Buffalo: An Economic and Public Health Concern

Water buffalo is an indispensable livestock in Asia and other countries due to its high meat and milk quality, aside from draft power source. It adapts well to tropical climate and has significant contribution to the livestock industry, provided with improved breeding and good animal husbandry practices. Infectious diseases are hindrance to good reproductive performance of livestock, resulting in huge economic loss. In addition, most of these diseases are zoonotic, posing serious threats on human health. However, its degree of severity varies in each region and is often overlooked. This chapter reviews the common and current updates on emerging bacterial, viral, protozoal, fungal and endoparasitic pathogens that infect water buffaloes worldwide. All of the diseases directly affect the animals’ health condition except for schistosomiasis where water buffalo played an important role as shedder of infection to humans. Leptospirosis, brucellosis, Bovine Tb, BVDV and fasciolosis have projected economic impact to water buffalo industry as well as its effect as zoonoses. However, the data seem underquantified since most are neglected diseases and are highly prevalent in developing countries. Further studies are needed particularly in countries where water buffalo is the major livestock than cattle to fully utilize the potential of the animal

TRYPANOSOMA EVANSI AND NEOSPORA CANINUM AMONG WATER BUFFALOES (BUBALUS BUBALIS) IN THE PHILIPPINES

The study determined the positivity rate of Trypanosoma evansi and Neospora caninum antibodies in water buffaloes in the province of Nueva Ecija, Philippines using Polymerase Chain Reaction (PCR) for T. evansi and competitive Enzyme-linked Immunosorbent Assay (cELISA) for N. caninum antibodies . A total of 100 whole blood and 100 serum samples were collected to test for T. evansi and N. caninum , respectively. Rotat 1.2 VSG gene was target using PCR for T. evansi detection. Neospora caninum antibody detection was done from the serum samples using cELISA test kit.Results revealed that the positivity rate of T. evansi in Nueva Ecija was 11% (11/100). The positive animals identified were from the municipalities of Muñoz (4/16; 25%), Sta. Rosa (3/13; 23.08%) and Talugtug (4/16; 25%). The seropositive rate of Nueva Ecija for N. caninum. was 46% (46/100), seropositive animals were identified in Cabanatuan City, 57.14% (4/7); Science City of Muñoz, 43.14% (22/51); Sta. Rosa, 40% (4/10); Sto. Sunday, 50% (6/12); and Talugtug 50% (10/20). The seropositivity rate of N. caninum and the presence of T. evansi in Nueva Ecija may contribute to the cases of abortions in the province and further studies should be employed to confirm the association of these organisms to abortion cases on water buffaloes

In vivo procjena učinka tripanocidnih lijekova na izolate protozoona Trypanosoma evansi izdvojene iz azijskih vodenih bivola (Bubalus bubalis).

The effects of the trypanocidal drugs against Trypanosoma evansi isolated from Philippine water buffaloes from the three island groups were comparatively evaluated. Specifically, the study determined the duration of the efficacy, relapse and death per drug dosage using laboratory mice. A total of 270 inbred Balb/c mice were divided into three groups corresponding to the three trypanosome isolates (Luzon, Visayas, and Mindanao). Each group had three sets, corresponding to the three trypanocidal drugs used with five treatment levels, and one control group each. Each experimental group was composed of five mice. Each mouse was with 0.2 mL of T. evansi intraperitoneally and blood was examined under the microscope. Parasitemia level was determined using the “Rapid Matching Method”. Effective and curative doses were noted and evaluated through t-test and bio-assay graphical analyses. Results showed that the Luzon isolate was sensitive to ≥5mg/kg of diminazene diaceturate and ≥10 mg/kg of both isometamidium chloride and quinapyramine sulphate + chloride. The Visayas isolate was sensitive to ≥5 mg/kg, ≥10 mg/kg, and ≥3 mg/kg of diminazene diaceturate, isometamidium chloride and quinapyramine sulphate + chloride, respectively. The Mindanao isolate was sensitive to ≥3 mg/kg of diminazene diaceturate and quinapyramine sulphate + chloride and 20 mg/kg of isometamidium chloride. The study suggested diminazene as the recommended drug against Luzon isolates, quinapyramine against Visayas isolates and either diminazene or quinapyramine against Mindanao isolatesIstraženi su učinci tripanocidnih lijekova na izolate vrste Trypanosoma evansi izdvojene iz triju otočnih skupina filipinskih vodenih bivola. Određivano je trajanje učinkovitosti, pojava recidiva i uginuća u laboratorijskih miševa. Ukupno je 270 miševa Balb/c bilo podijeljeno u tri skupine što je bilo sukladno trima izolatima s tri filipinska otoka i to Luzon, Visayas i Mindanao. Svaka skupina bila je podijeljena u tri podskupine koje su peterokratno dobivale tri različita lijeka. Unutar svake skupine određena je i kontrolna podskupina. U svakoj pokusnoj skupini bilo je pet miševa. Svakom mišu bila su intraperitonejski ubrizgana 0,2 mL kulture T. evansi i izvađena krv za mikroskopsku pretragu. Razina parazitemije određivana je brzom metodom (engl. rapid matching method). Učinkovitost i ljekovita doza vrednovane su metodom (engl. rapid matching method). Učinkovitost i ljekovita doza vrednovane su t-testom i grafičkom analizom biološkog pokusa. Rezultati su pokazali da je izolat Luzon bio osjetljiv na ≥5 mg/kg diminazenova diaceturata i ≥10 mg/kg izometamidijeva klorida i kombinacije kvinapiramin sulfata i klorida. Izolat Visayas bio je osjetljiv na ≥5 mg/kg diminazen diaceturata, ≥10 mg/kg izometamidij klorida i ≥3 mg/kg kombinacije kvinapiramin sulfata i klorida. Izolat Mindanao je bio osjetljiv na ≥3 mg/kg diminazen diaceturata i kombinacije kvinapiramin sulfata i klorida, te 20 mg/kg izometamidij klorida. Može se reći da je diminazen bio učinkovit na izolate s otoka Luzon, kvinapiramin na one s otoka Visayas, dok su diminazen ili kvinapiramin bili učinkoviti na izolate s otoka Mindanao

Screening of BCL-2 associated X protein gene polymorphism associated with scrotal hernia in domesticated swine using polymerase chain reaction-restriction fragment length polymorphism

Objective This study was conducted to screen scrotal hernia in domesticated swine from selected breeders in the Philippines. This defect is associated with a cytosine to thymine mutation in the BCL-2 associated X protein (BAX) gene of swine. Methods Genetic screening was done by DNA extraction followed by amplification and digestion using polymerase chain reaction-restriction fragment length polymorphism, amplifying the 416 bp region of the BAX gene that was subjected to digestion using the Ear I enzyme. Sequencing was also conducted to validate the results. Results Results revealed that out of 538 samples tested, 411 (76.4%) of the samples were found to be normal whereas the remaining were carriers of the mutation in which 80 (14.9%) were heterozygous mutants and 47 (8.7%) were homozygous mutants. Pietrain breed was found to have the highest incidence. Conclusion Having a scrotal hernia eliminates the chances of using the boar as a breeder stock because the following generations arising from it would most likely exhibit herniation. It is therefore advised to establish a genetic screening method for Scrotal Hernia in the Philippines to eliminate the negative gene from the herd

Application of Noble Metals in the Advances in Animal Disease Diagnostics

The advent of molecular biology and biotechnology has given ease and comfort for the screening and detection of different animal diseases caused by bacterial, viral, and fungal pathogens. Furthermore, detection of antibiotics and its residues has advanced in recent years. However, most of the process of animal disease diagnostics is still confined in the laboratory. The next step to conduct surveillance and prevent the spread of animal infectious diseases is to detect these diseases in the field. Through the discovery and continuous development in the field of nanobiotechnology, it was found that incorporation of noble metal nanoparticles to biotechnology tools such as the loop-mediated isothermal amplification (LAMP), lateral flow assays (LFAs) and dipsticks provided a promising start to conduct point-of-care diagnostics. Moreover, the modification and application of nanoparticle noble metals has increased the stability, effectiveness, sensitivity and overall efficacy of these diagnostic tools. Thus, recent advances in disease diagnostics used these noble metals such as gold, silver and platinum

Sensitive detection of Mycobacterium bovis in spiked milk using polymerase chain reaction assay

Bovine tuberculosis is a chronic zoonotic disease that affects both animal and human health and imposes serious public health concerns in the world. Intake of non-pasteurized milk is considered the most probable vehicle for the transmission of pathogenic bacteria. In this study, the detection of Mycobacterium bovis BCG in spiked milk using a polymerase chain reaction was performed. The performance of two DNA extraction methods, CTAB/phenol:chloroform:isoamyl alcohol and EXTRAGENMB were also evaluated. In addition, Mycobacterial concentration was tried to determine using the Standard/ Viable Plate Count Method and Spectrophotometric (Turbidimetric) Method. PCR successfully detected M. bovis BCG in spiked milk, detecting approximately up to two bacilli per reaction. The two DNA extraction methods were effective in the isolation of amplifiable DNA, having the advantage of EXTRAGENMB in terms of (1) shorter duration of DNA extraction, (2) less sample manipulation, and (3) ease of execution of the procedure. Quantitative determination of the Mycobacterial population however failed to quantify the bacterial concentration per dilution, suggesting that CFU concentration should be considered an approximation. It is expected that this method can be used for the detection of M. bovis in raw milk samples

Molecular characterization of the lymphocyte activation gene-3 (LAG-3, CD223) of swamp- and riverine-type water buffaloes (Bubalus bubalis)

The present study was conducted to characterize LAG-3 of swamp- and riverine-type water buffaloes by DNA sequencing, homology and phylogenetic analysis. Bubaline LAG-3 sequence contained an open reading frame of 1551 nucleotide, encoding a polypeptide of 516 amino acids. Nucleotide and amino acid sequence homology of LAG-3 revealed 76-96% and 61-94% identity in water buffalo to that of other mammals, respectively. LAG-3 protein sequence of water buffalo contained four extracellular domains, a transmembrane domain and different conserved regions. There were three N-glycosylation sites, two sequence motifs: ‘RGD’ and ‘WXC’ motif and five cysteine residues located at different positions of extracellular region. Likewise, the possible serine phosphorylation site and the ‘KTGELE’ inhibitory motif were found in the intracellular region of bubaline LAG-3. However, one highly conserved cysteine residue in mammalian LAG-3 was replaced by tyrosine in both swamp- and riverine-type water buffaloes. Phylogenetic analysis generated high bootstrap value between the two types of water buffalo which further confirmed the degree of relationship between bubaline species. This was the first report that describe the genetic characteristic of LAG-3 in swamp- and riverine-type water bufffaloes

Biofilm: a platform for the evaluation of antimicrobial treatments and prevention against Mycoplasma bovis infection

Mycoplasma bovis is an important pathogen in large ruminants that causes various clinical pathologies with great economic impact. Though mycoplasmas were thought to have gone reductive evolution that led to the loss of genetic materials leading to the marked absence of cell wall and canonical virulence factors, adaptation made by some M. bovis isolates include production of biofilms to survive, evade host immune response, persists in a biotic environment and may mount related responses that render chemotherapy ineffective. In this review, topics on biofilms commonly produced by M. bovis isolates were discussed to view its pathogenicity. Reports that describe the involvement of variable surface antigensin biofilm production of M. bovis are also presented. This information invites interest on biofilms anchored on studies where it can be developed to be utilized as platforms for the evaluation of antimicrobial treatments and for the identification of a preventive and control strategy against Mycoplasma bovis infection