RECENT DEVELOPMENT & CONTROL OF BOVINE MASTITIS-2.pptx

RECENT DEVELOPMENT & CONTROL OF BOVINE MASTITIS-2.pptx

1. RECENT DEVELOPMENT IN DIAGNOSIS OF BOVINE MASTITIS KAINAT SHEHZADI 2023-MPHIL-1111 DISEASES OF MAMMARY GLAND

2. Recet Developement in Bovine Mastitis Bovine mastitis remains a significant threat to dairy production worldwide, affecting 30-40 % cows annually milk quality, animal welfare, and farm profitability. This complex and multifactorial disease results in significant economic losses, estimated at over $2 billion annually in the United States alone, due to reduced milk production, discarded milk and treatment cost. Despite advances in management and treatment, mastitis continues to be a major challenge for the dairy industry.

3. Recet Developement in Bovine Mastitis Conventional diagnostic tests for mastitis are usually qualitative with lesser specificity and sensitivity, whereas advanced tests are quantitative, highly specific and sensitive (Godden et al. Citation2017; Hussein et al. Citation2018; Chakraborty et al. Citation2019).

4. Advance/ Recent Tests Conventional Diagnostic Tests PCR (Multiplex/ Real time) Nanotechnology Stem cell technology Micro array technology Laser therapy Bio-chips Detection of bio marker including heptoglobin, Serum amlyoid California Mastitis Test (CMT) R-mastitest Mast-O-test Somatic cell count Bulk Tank Somatic Cell Count (BTSCC) Enzymes (N-acetyl-D glucosaminidase; lactate dehydrogenase) pH indicator Recet Developement in Bovine Mastitis

5. Nucleic acid base diagnosis: Molecular assays became the ideal standard for mastitis diagnosis in the last decade.They allow rapid, quantitative, qualitative, and wide-scale diagnosis. These methods are based on nucleic acid detection, such as polymerase chain reaction (PCR). Different types of PCR are used to identify the genome structures of the pathogens causing mastitis: amplification of their DNA fragment for their detection only (conventional PCR), detection and quantification of one pathogen (reverse transcription polymerase chain reaction [RT-PCR]), and detection and quantification of the various pathogens in the same sample (multiplex PCR).

6. Nucleic acid based diagnosis: The first two techniques for mastitis detection are conventional PCR and multiplex PCR . Both methods have primers designed for 23S and 16S ribosomal ribonucleic acid (rRNA). Gillespie and Oliver used multiplex PCR to detect S. aureus, Str. uberis, and Str. agalactiae instantaneously using primers rather than 16S and 23S rRNA. Multiplex PCR was first designed to detect four pathogens in a single test and was expanded to detect nine or 11 pathogens in a single reaction. Shome et al. used multiplex PCR to identify ten types of bacteria that cause bovine mastitis simultaneously.

7. Advantages: These tests have several merits, such as high Se and Sp, speed, and avoidance of the drawbacks related to culture-based tests. Moreover, they can detect the bacteria in milk samples even in the presence of drug residues and preservatives. PCR products can be stored in a refrignator or freezer for an extended period.

8. Advantages: Compared to the polymerase chain reaction (PCR), culture plate methods were able to identify mastitis pathogens on only 47% of the no-growth milk samples. Nucleic acid-based detection or PCR has facilitated the detection of those pathogens that cannot be identified using standard bacterial culture plates.

9. Disadvantages: Limited access in developing coutries. False-postive results can be found when milk residues are in the milking machines, contamination and colonizationof the teat canal. They can also arise when using primers that are not sufficiently specfic or when the cycle conditions are changed. In addition clotted milk samples or the presence of PCR inhibitors in the milk samples can result in incorrect DNA extraction and purification, which can lead to a false-negative result. On the other hand, this issue can be overcome by column purification.

10. Microarray technology: Microarray technology depends on hybridizing different types of target genes overloaded on microarray chips. It imagines them after exposure to the complementary DNA (cDNA) probe bound with chemiluminescent or fluorescence stains. Vidic et al. reported that more than seven pathogens-causing mastitis could be detected in a single reaction by developing multiplex biochips. Lee et al. showed that six field strains for four pathogens can be identified and evaluated in a single milk sample in less than three hours through PCR and a nucleic acid microarray immunoassay.

11. Proteomic methods: Proteomic methods can analyze high-throughput proteins, detect and specify them, and predict the changes in protein levels, resulting in a comprehensive understanding of the occurrence of mastitis and the progression of the disease. On the other hand, all progress in proteomics and these technological techniques are expensive and not generally used for routine mastitis diagnosis in farm animals.

12. Proteomic methods: In addition, they have limitations in their validation procedures because of the lack of species-specific antibodies. Furthermore, there is no distinctive protocol for analyzing many less abundant milk proteins representing 5% of the total milk proteins in the whey or the milk fat globule membrane.

13. Proteomic methods:

14. Detection of bio markers: APPs has become an important diagnostic indicator used in human and animal medicine. Haptoglobin (Hp) is one of the most abundant APPs produced by the liver, and its concentration is increased in serum because of bacterial inflammation and infection. In addition, it is used to differentiate between SCM and CM and between healthy and diseased cows. Hp is significantly elevated in milk during mastitis. Georgieva et al. suggested that the concentration of plasma Hp in the high SCC group (SCM) was approximately six times higher than that in the normal SCC group.

15. Detection of bio markers: Serum amyloid A (SAA), which is synthesized in the liver or extrahepatic origins as in the mammary gland, is another pattern of APP. SAA is increased 1,000 times within 24 h of tissue injury because this protein has pro-inflammatory and anti-inflammatory effects. Hence, it can be used to monitor the health status and prognosis of veterinary medicine therapy. The level of this protein is higher in mastitic milk than in normal milk, followed by an increase in SCC.

16. Detection of bio markers:

17. Novel techniques: Novel techniques for detecting early udder inflammation within hours after infections are under development. For example, transrectal color Doppler sonography is a noninvasive technique that detects the blood flow volume that passes to the mammary gland within a few hours after the infusion of a microorganism. Considering that the amount of blood flow passing to the udder changes after infection, the disease can be detected in the early stages before severe losses occur.

18. Detection of bio markers: In addition, it is higher in the inflamed quarter than in the normal quarter of the same udder. Eckersall et al. reported a significant correlation between Hp in milk and serum. On the other hand, there was no relationship between the concentration of SAA in milk and serum.

19. Cytometric Fingerprinting Machine Learning(CFML): Cytometric fingerprints largely resembled SCC technique, CFML toolchain as a label-free, objective, high-throughput microbiological milk quality evaluation method for routine mastitis screening. Flow cytometry is used to detect t he presence, number and distribution of somatic cells (shed epithelial cells, leukocytes - macrophages, polymorphonuclear neutrophils-PMN, lymphocytes), hence infer sub-clinical mastitis at an early stage It is used to identify the lactation stage and pathogen intensity.

20. Cytometric Fingerprinting:

21. B-mode ultrasonography B-mode ultrasonography was also used to determine the disturbance in milk secretion and the structural changes in the teat cistern and udder tissue. For the first time in mastitis, Risvanli et al. evaluated the supramammary lymph nodes of cattle by color Doppler ultrasonography because mastitis causes lymphocytic proliferation that leads to morphological changes in the supramammary lymph nodes.

22. m

23. Infrared thermography (IRT): Infrared thermography (IRT) is a noninvasive, rapid, safe, and sensitive method to detect the early inflammatory changes accompanied by SCM by measuring the changes in the teat skin temperature. IRT is sufficient alone to diagnose mastitis and detect any changes that have not yet been observed in cows. The Se and Sp of IRT for the diagnosis of SCM are similar to those of CMT. Pezeshki et al. reported a 2–3°C increase in udder temperature in cows infected with E. coli compared to the control.

24. Infrared thermography (IRT): This result is similar to Metzner et al. who reported that the temperature of the dairy cow udder was 2.06°C higher in cases of acute mastitis caused by E. coli infusion than in normal cows. Holstein Friesian cows infected with SCM showed increases in udder temperature of 2–3°C, 0.72–1.05°C , or 1.35°C on the surface of the udder between infected and healthy quarters.

25. Other molecular techniques: Other molecular techniques include real-time quantitative PCR (qPCR), loop-mediated isothermal amplification (LAMP), and next-generation sequencing (NGS) methods. As a secondary confirmatory test, matrix-assisted laser desorption/ionization time-of-flight (MALDI–TOF) mass spectrometry can be applied as a diagnostic technique for bacterial species identification in mastitis studies

26. REFERENCES: Bovine mastitis: risk factors, therapeutic strategies, and alternative treatments — A review Basic concepts, recent advances, and future perspectives in the diagnosis of bovine mastitis Advances in therapeutic and managemental approaches of bovine mastitis: a comprehensive review Cytometric fingerprinting and machine learning (CFML): A novel label-free, objective method for routine mastitis screening