ABOUT OUR TECHNIQUES

Each panel is specifically designed according to the species, the clinical picture, the age and the production stage, providing a more complete differential diagnosis adapted to the infectious diseases that affect production animals.

A more complete diagnosis

We study at the same time the main infectious agents associated with a single clinical process, helping to identify both single infections and co-infections.

Adapted to each case

Panels are designed taking into account the species, the age or production stage and the clinical process, selecting the most relevant pathogens in each situation.

Optimised costs and samples

From a single sample we can analyse different pathogens by qPCR, optimising the use of the material received and reducing the cost compared with running multiple independent tests.

Fast results

The automation of the process and the analysis by real-time PCR make it possible to obtain results quickly, supporting decision-making on the farm.

This information is particularly useful for carrying out epidemiological studies, determining genotypes, identifying the most relevant variants and serotypes, differentiating strains and analysing the relationship between microorganisms from different animals, farms or outbreaks.

Characterise viruses and bacteria

We identify genotypes, variants, serotypes and other genetic markers that make it possible to characterise with greater precision the microorganisms present in a sample.

Study the relationship between strains

Comparing sequences and genetic profiles makes it possible to assess the degree of similarity between microorganisms isolated from different animals, batches, farms or clinical episodes.

Carry out epidemiological studies

Molecular characterisation helps to investigate the circulation of infectious agents, to establish possible relationships between outbreaks and to study the traceability of the strains present on a farm or in an animal population.

Type bacterial strains

We apply techniques such as MLST (Multilocus Sequence Typing) and MLVA (Multiple-Locus Variable-number Tandem Repeat Analysis) to differentiate and classify bacterial strains according to their genetic profiles.

Depending on the diagnostic objective, we use Sanger sequencing, nanopore sequencing and molecular typing methods, complemented with qPCR when the targeted identification of specific genotypes or variants is required.

Sanger sequencing

It provides the sequence of specific regions of the genome with high accuracy. It is especially useful for the identification and characterisation of defined regions of the genome of microorganisms, the confirmation of variants and the study of specific genes.

Third-generation sequencing using nanopores

Nanopore sequencing technology provides DNA or RNA sequences that support characterisation and molecular epidemiology studies.

After extracting the DNA or RNA from the sample, a specific region of the pathogen genome is amplified, which makes it possible to detect the presence of the agent in real time with high sensitivity and specificity.

In pathogens whose genetic material is RNA, a reverse transcription is carried out beforehand, giving rise to the RT-qPCR technique (Reverse Transcription quantitative PCR).

Detection and identification

qPCR makes it possible to detect specifically the genetic material of a pathogen even when it is present in small amounts.

As it works directly on the sample, it supports a rapid diagnosis of infectious diseases without depending on the previous isolation or culture of the microorganism.

Estimation of the pathogen load

The Cq value (Quantification Cycle), also called Ct (Cycle Threshold or Threshold Cycle), corresponds to the amplification cycle at which the fluorescent signal exceeds the threshold set by the assay.

In general terms, the lower the Cq/Ct value, the greater the amount of genetic material of the pathogen present in the sample.

This value can provide additional information to interpret the result within the clinical and epidemiological context, always taking into account the type of sample, the pathogen analysed and the characteristics of the assay.

Molecular characterisation and typing

Using specific qPCR assays we can detect virulence genes and toxins, and identify serotypes, genotypes, mutations or specific variants.

This characterisation provides immediate information about the agent detected and can help to guide prevention and control measures, as well as the selection and characterisation of isolates intended for further studies.

Differentiation between vaccine and field strains

When specific genetic markers exist, we can use qPCR assays to differentiate vaccine strains from field strains.

This information is especially useful in vaccination programmes, epidemiological investigations and the interpretation of positive results after certain vaccines have been administered.

1. Sample preparation

We select and process the appropriate sample according to the pathogen and the clinical process we want to investigate.

2. Nucleic acid extraction

We extract and purify the DNA or RNA present in the sample.

3. Specific amplification

We use primers and probes designed to recognise specific regions of the pathogen genome.

4. Real-time detection

The instrument records the fluorescence generated during each amplification cycle and determines the Cq/Ct value.

5. Interpretation of the result

The result is interpreted together with the other results, taking into account the controls of the assay, the Cq/Ct value, the type of sample and the clinical and epidemiological context.

• Confirm the presence and viability of a bacterium involved in the clinical process.
• Obtain the bacterial strain in order to carry out further studies.
• Select the bacterial isolates intended for the production of autogenous vaccines.
• Carry out antimicrobial susceptibility studies.
• Keep strains of interest for R&D.

Sample selection

We select and process the most suitable samples according to the animal species, the clinical process and the bacteria we want to investigate.

Culture in specific media

We seed the samples in culture media adapted to the requirements of each bacterium, including specific conditions for the growth of aerobic and anaerobic bacteria.

When necessary, we use pre-enrichment media or steps that favour the recovery of bacteria present in low concentrations or with special growth requirements.

Bacterial identification

Once a pure culture has been obtained from a single isolated colony, we identify it by MALDI-TOF mass spectrometry.

MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry) is a mass spectrometry technique that makes it possible to identify bacteria from their characteristic protein profile.

A small amount of the isolated bacterial strain is placed on the plate of the instrument. The MALDI-TOF generates the characteristic mass spectrum of the microorganism and compares it automatically with a large reference database in order to determine its identity.

In most cases, this technology makes it possible to reach an identification at genus and species level.

Rapid identification

Once the bacterium has been isolated, identification by MALDI-TOF can be carried out in a few minutes.

High identification capacity

It makes it possible to identify numerous bacterial species of veterinary interest using a single technology.

Precise and objective identification

Comparing the protein profile with reference spectra provides a reliable identification when the microorganism is correctly represented in the database.

It supports further studies

A precise identification of the isolate makes it possible to select the most suitable complementary tests, such as antimicrobial susceptibility studies, molecular characterisation or the selection of strains for autogenous vaccines.

It is a particularly useful tool in livestock farming to find out about previous contact with certain pathogens, to study the circulation of diseases on a farm and to monitor the health of animals, batches or herds.

We work with reference commercial kits and validated immunological techniques, selected according to the animal species and the agent to be investigated.

Antibody detection

It makes it possible to identify specific antibodies against different infectious agents and to know whether the animal has been in contact with these pathogens and has developed an immune response against them.

Herd monitoring

Analysing individual animals or groups makes it possible to study the seroprevalence and the circulation of pathogens within a farm, supporting the follow-up of health programmes.

Serological profiles

In certain cases we can design serological profiles adapted to the objective of the study, selecting representative animals, batches or age groups.

These profiles make it possible to investigate specific situations, to compare different groups within a farm and to study the serological evolution of a herd over time, providing useful information to assess changes in exposure, the circulation of infectious agents or the response to control and vaccination programmes.

Assessment of vaccination programmes

In certain diseases, serology can be used to assess the immune response after vaccination and to monitor immunity at batch or herd level.

Epidemiological studies

Serological profiles help to find out the exposure of a population to certain infectious agents and to study its evolution over time.

ELISA

The enzyme-linked immunosorbent assay (ELISA) makes it possible to detect and, depending on the assay, to estimate the amount of specific antibodies present in the sample.

It is a technique widely used in veterinary diagnostics for its sensitivity, reproducibility and capacity to analyse a large number of samples.

Agglutination

Agglutination techniques detect the reaction between specific antibodies and their corresponding antigens, producing a visible agglutination that makes it possible to determine the presence of antibodies against the agent studied.

Immunofluorescence

Immunofluorescence makes it possible to detect antibodies through their specific binding to antigens and their visualisation using fluorescent markers.

The presence of antibodies indicates that the immune system has been in contact with a given antigen, but it does not always mean that there is an active infection at the time of the analysis.

The interpretation must be carried out taking into account factors such as the age of the animal, the vaccination status, the time of sampling, the presence of maternal antibodies, the epidemiology of the farm and the characteristics of each disease.

When necessary, serology can be complemented with direct diagnostic techniques such as qPCR or microbiological culture in order to obtain a more complete view of the clinical process.

Using high-performance liquid chromatography techniques (HPLC and UHPLC) we can investigate suspected poisoning and analyse compounds of toxicological interest in complex matrices.

Toxicological analyses

We carry out analyses aimed at the detection, identification and quantification of toxic substances when poisoning is suspected.

The choice of the analysis and of the most suitable sample depends on the suspected substance, the animal species, the clinical signs and the circumstances of the case.

Mycotoxins

We analyse mycotoxins of relevance in animal health and production. Mycotoxins are toxic compounds produced by certain fungi that can contaminate raw materials and feed.

Their detection and quantification help to assess the possible exposure of the animals and its relationship with health or production problems.

HPLC (High-Performance Liquid Chromatography) is an analytical technique that makes it possible to separate the different compounds present in a sample in order to then detect, identify and quantify them.

1. Sample preparation

The sample is processed in order to extract the compounds of interest and to reduce possible interferences from the matrix.

2. Chromatographic separation

The sample is introduced into the chromatographic system, where its components are separated according to their physicochemical properties and their interaction with the mobile and stationary phases.

3. Detection and identification

The separated compounds are detected by the corresponding analytical system and are identified using reference parameters and standards.

4. Quantification

When the analysis allows it, we determine the concentration of the compound detected in order to assess the level of exposure present in the sample.

UHPLC (Ultra-High-Performance Liquid Chromatography) is an evolution of high-performance liquid chromatography that works with systems capable of offering a more efficient chromatographic separation.

This technology provides greater resolution, sensitivity and speed of analysis, supporting the detection and quantification of compounds present in complex matrices and at low concentrations.

In addition to their usefulness in individual clinical cases, in production animals they are a very useful tool to monitor batches and herds, to detect subclinical alterations and to study their relationship with management, feeding and productive performance.

Assessment of health status

They make it possible to assess the general condition of the animals and to detect alterations that may be related to infectious, inflammatory, metabolic or nutritional processes.

Detection of subclinical processes

Certain haematological or biochemical changes may appear before evident clinical signs exist, helping to identify health problems at early stages.

Follow-up of batches and herds

Analysing representative groups makes it possible to study the evolution of a population and to relate certain parameters to its health status and productive performance.

Assessment of changes and treatments

Blood profiles can be used to assess the evolution of the animals after a treatment or after introducing changes in management, feeding or farm conditions.

Comparative studies

They make it possible to compare groups subjected to different diets, additives, management practices, environmental conditions or genetic lines, providing objective indicators of their physiological response.

BIOCHEMISTRY

Blood biochemistry makes it possible to measure different substances present mainly in serum or plasma and to obtain information about the functioning of different organs and metabolic processes. Among other parameters, we can assess:

• Metabolites and excretion products, related to different metabolic processes and to the function of certain organs.
• Enzymes, whose variations may be associated with cell and tissue injury or alteration.
• Proteins and other blood components, related to the metabolic, nutritional or inflammatory status.
• Nutrients and minerals, useful to assess the nutritional status and certain metabolic imbalances.

The selection of the parameters can be adapted to the clinical, productive or experimental objective of each case.

HAEMATOLOGY

Veterinary haematology studies the cells present in the blood and their main characteristics by means of a blood count.

Red cell line

We assess the erythrocytes or red blood cells and the parameters related to them, providing useful information to detect and characterise alterations such as anaemia or changes in the concentration of blood cells.

White cell line

We study the leukocytes or white blood cells, both their total number and their distribution, providing information about the immune response and about possible inflammatory or infectious processes.

Platelet line

We assess the platelets and the associated parameters, which are essential in the mechanisms of haemostasis and coagulation.

The results must be interpreted taking into account the species, age, production stage, physiological status, feeding and clinical situation of the animals.

In farm studies, assessing several animals together and their evolution over time can provide more information than an isolated result, helping to detect trends and to compare groups within the same population.