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INGUBAL Ruminant Lechero®. Postbiotics for mastitis prevention in dairy cattle.
Ingubal Ruminant Lechero. INGULADOS

This article presents a groundbreaking study that designs and validates INGUBAL Ruminant Lechero®  as a postbiotic to address mastitis in dairy sheep. The study design included the characterization of pathogenic and beneficial bacterial strains, highlighting the selection of lactic acid bacteria with antimicrobial and immunomodulatory properties. Postbiotic formulation INGUBAL Ruminant Lechero®  from the fermentation of three bacterial strains from the BAL-INGULATES collection was presented as a complementary feed that includes by-products of bacterial metabolism and inanimate microbial cells. During the supplementation period, a significant reduction of 16.77% in somatic cell count was observed, indicating an improvement in mammary health and milk quality. The presence of the pathogenic bacterium S. aureus was completely eliminated after 60 days of administration, being a frequent cause of subclinical mastitis affecting dairy production. In addition, the capacity of the postbiotic is highlighted. INGUBAL Ruminant Lechero® to modulate the intestinal microbiota, suggesting a comprehensive and sustainable approach for the treatment and prevention of mastitis in dairy production. These results offer promising prospects toward more effective and environmentally friendly alternatives in dairy mammary health management.

 

INTRODUCTION

The term mastitis can be defined as a process that involves inflammation of the mammary gland tissues, which can affect both the quality and quantity of milk production. This condition, when of infectious origin, is caused by various pathogens that invade the udder, giving rise to a series of health problems in dairy ruminant farms, which have a great economic impact causing greater losses than the rest of the existing pathologies in the farm as a whole. Among the most frequent pathogens that play a prominent role in the development of mastitis are bacteria such as Staphylococcus aureus, Streptococcus agalactiae, Escherichia coli and other microorganisms. Given the wide variety of aspects that can characterize mastitis, different categories have been formulated to address this condition. Focusing on the clinical signs that affected animals may show, we find:

  • Subclinical mastitis: these are processes in which the inflammation in the udder does not manifest any clinical signs evident to the naked eye. The only observable manifestation is reflected in the increase in somatic cell count in milk and the decrease in milk yield.
  • Clinical mastitis: refers to those that generate noticeable clinical signs, both locally, specifically in the udder, and sometimes systemically, such as fever or lack of appetite. Depending on the evolution of the process, a distinction can be made between acute, subacute and chronic mastitis.

To deal with mastitis of infectious origin, it is essential to focus on treating affected animals and applying preventive measures to avoid the onset of the disease. Historically, both treatment and prevention have been based on the use of antibiotics, either systemic or intramammary, both in affected animals and in those at risk as a preventive measure. However, the indiscriminate use of antibiotics over time has led to the emergence of bacterial resistance, threatening the efficacy of these drugs and posing risks to public health. Therefore, the need to optimize the use of antibiotics demands an evolution in these traditional practices, as well as the search for more sustainable alternatives.

In this sense, postbiotics emerge as a promising alternative in the treatment and prevention of mastitis. Postbiotics are preparations of inanimate beneficial microorganisms and metabolites resulting from their fermentation that have positive health effects due to their antimicrobial and immunomodulatory properties. Postbiotics can act by modulating the gut microbiota and there is increasing scientific evidence of the impact this may have on the mammary microbiota. In addition, by modulating the inflammatory response at the systemic level, postbiotics not only fight the current infection, but also help prevent future episodes of mastitis. Unlike traditional antibiotics, INGUBAL Ruminant Lechero® offers a more sustainable option, reducing the risk of bacterial resistance and minimizing environmental impacts. Its focus on long-term health represents a significant advance in the sustainable management of mammary health in dairy production.

 

METHOD

1. Product design: selection of beneficial bacteria

The first step in designing a postbiotic is the characterization of bacterial strains, both pathogenic and beneficial. The product design is a combination of a complete clinical history with comprehensive veterinary diagnostics along with in vitro microbiological testing for strains producing molecules with antimicrobial and immunomodulatory properties.

1.1. Pathogen isolation

Data were collected from medical records in a Lacaune dairy sheep farm and the microorganism Staphylococcus aureus, a Gram-positive bacterium that causes mastitis in females, was isolated.

1.2. Selection of beneficial bacteria

INGULADOS has a collection of bacterial strains that have been isolated from animals in exceptional health and have the ability to produce antimicrobial and immunomodulatory substances. Based on our previous results, we made a selection of lactic acid bacteria with the ability to inhibit the growth of S. aureus and which also possess molecules that regulate the inflammatory response at the systemic level.

2. Product development: manufacturing and validation

2.1. Product manufacturing

Based on the microbiological characterization and the information obtained from the previous phases, a combination of bacteria and their metabolites was selected to elaborate the final product to favor mastitis control. Fermentation was performed under optimal conditions for each strain chosen to produce the maximum amount of antimicrobial metabolites and immunomodulatory molecules.

The final product was named INGUBAL Ruminant Lechero® and is a complementary feed consisting of a postbiotic that includes the by-products of beneficial bacteria metabolism as well as the inanimate microbial cells themselves.

2.2. Product validation

The final step of validation of the effectiveness of the product was carried out in a herd with 840 Lacaune dairy sheep, to which the following was added INGUBAL Ruminant Lechero®. to the mothers’ feed at a concentration of 0.2% (2 kg per Tn of feed) during the entire milking period.

Livestock productivity data were collected by somatic cell count per milliliter, and the results were compared with data obtained in the same period in previous years and microbiological study was performed to detect the presence of S. aureus before and after supplementation.

 

RESULTS AND DISCUSSION

The results obtained in the study provide promising evidence for the efficacy of the postbiotic INGUBAL Ruminant Lechero® in the control of mastitis in Lacaune dairy sheep.

Eighty-two milk samples were collected during the supplementation period and the mean somatic cell count was 933 celsx1000/mL ± 129.55. This data was compared with the same period of the previous year (1121 ± 138.97 celsx1000/mL, n=82), obtaining statistically significant differences (t test: <0.0001) with a reduction of 16.77%. The significant reduction in somatic cell count during the supplementation period indicates an improvement in the mammary health of the ewes, which translates directly into higher quality milk produced. This finding is especially relevant, since subclinical mastitis, even without obvious clinical signs, negatively affects milk production.

Results of somatic cell counts in supplementation periods. INGULATED.

Figure 1 – Results of somatic cell counts in the supplementation periods.

S. aureus bacteria were detected in 50% of the samples before supplementation, at a mean concentration of 1.21-104 Colony Forming Units per milliliter (cfu/mL). From 60 days after the beginning of the administration of INGUBAL Ruminant Lechero® and until the end of the study, the presence of pathogenic bacteria was not detected in any of the samples. This 100% reduction percentage suggests an effective action of the postbiotic in the inhibition of this Gram-positive mastitis-causing bacterium. This result is particularly noteworthy given the importance of S. aureus in the generation of mastitis and its negative impact on milk production and quality. Although it is sometimes associated with subclinical infections, it has a great impact on productivity and milk quality, increasing somatic cell counts.

Isolation and identification of Staphylococcus aureus in milk. INGULATES

Figure 2 – Isolation and identification of Staphylococcus aureus in milk.

The metabolites and antimicrobial molecules produced by the beneficial bacteria in the INGUBAL Ruminant Lechero® postbiotic include bacteriocins, organic acids and others, and have been shown to have positive effects on the intestinal microbiota by promoting a more balanced and pathogen-resistant environment. Modulation of the gut microbiota may have direct repercussions on the mammary microbiota, as it is believed that there is a bidirectional communication between these two ecosystems, known as the gut-mammary axis, and is an emerging phenomenon in research on mammary health in dairy ruminants. On the other hand, the improvement in intestinal health may contribute to strengthening the immune system’s defenses at the systemic level which, together with the immunomodulatory molecules contained in the product, may reduce the predisposition to mammary infections.

 

CONCLUSIONS

Mastitis are inflammatory processes of the udder that are accompanied by a quantitative and qualitative alteration of milk production. These pathologies are of great importance in dairy cattle farms, causing greater losses than the rest of the pathologies as a whole. The bacterium Staphylococcus aureus causes contagious mastitis in sheep in a large number of cases, causing acute infection, although in many cases the presence of subclinical infections without symptoms is detected.

INGUBAL Ruminant Lechero® has been specifically designed to help control mastitis caused by S. aureus, because it contains bacterial strains with the capacity to inhibit the growth of the pathogenic bacteria, as well as immunomodulatory elements that help control inflammation. INGUBAL Ruminant Lechero® is an innovative post-biotic manufactured by
INGULATES
containing three inanimate bacteria from the BAL-INGULADOS collection in addition to their antimicrobial and immunomodulatory metabolites. Management of INGUBAL Ruminant Lechero® during the milking period is an affordable and sustainable strategy that can improve milk quality and help prevent complications associated with mastitis of infectious origin, especially subclinical mastitis.

 

REFERENCES

  • Bravo, Maria, et al.“Wildlife Symbiotic Bacteria Are Indicators of the Health Status of the Host and Its Ecosystem“. Applied and Environmental Microbiology 88.1 (2022): e01385-21.
  • Esteban-Blanco, Cristina, et al.“Microbiota characterization of sheep milk and its association with somatic cell count using 16s rRNA gene sequencing“. Journal of Animal Breeding and Genetics 137.1 (2020): 73-83.
  • Gussmann, Maya, et al.“Economic and epidemiological impact of different intervention strategies for subclinical and clinical mastitis“. Preventive veterinary medicine 166 (2019): 78-85.
  • Hu, Xiaoyu, et al.“The rumen microbiota contributes to the development of mastitis in dairy cows“. Microbiology spectrum 10.1 (2022): e02512-21.

Hu, Xiaoyu, et al.“Targeting gut microbiota as a possible therapy for mastitis“. European Journal of Clinical Microbiology & Infectious Diseases 38 (2019): 1409-1423.

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