Subcutaneous application of hyperimmune serum against Histophilus somni recombinant proteins affects serum antibody reactivity in beef calves.

BACKGROUND: Respiratory tract diseases cause significant economic loss in beef cattle. This study aimed to determine whether the application of hyperimmune serum (HS) containing antibodies against selected antigens of Gram-negative bacteria would improve the health and growth of different breeds of beef calves kept on three farms. Two recombinant protein antigens (Histophilus somni rHsp60 and rOMP40) were used to immunize four cows to produce HS. Eighty seven beef calves (Charolaise n = 36, Limousine n = 34, and crossbreed n = 17) were included into study. One hundred milliliters of serum were administered subcutaneously to 43 beef calves (Charolaise n = 18, Limousine n = 17, and crossbreed n = 8) twice, between 1 and 5 and 21-28 days of life. Calves were examined three times, and blood samples were taken to evaluate immunoglobulin M, G1, and G2, fibrinogen, serum amyloid A, and haptoglobin concentrations and reactivity of these Ig classes of antibodies against H. somni rHsp60 and rOMP40. Average daily weight gain during the first month and until weaning was calculated. RESULTS: HS showed higher (p ≤ 0.05) reactivity in calf sera against H. somni rHsp60 and OMP40 in IgG1 and IgG2. In experimental calves, compared to control calves, the reactivity of IgG1 against rOMP40 in the second sampling was higher in Limousine calves (p ≤ 0.001) and in the other two herds (p ≤ 0.05). Serum IgG2 antibody activity against H. somni rHsp60 in the second sampling was higher in experimental calves than in control calves in charolaise (p ≤ 0.05) and limousine (p ≤ 0.001) herds. The reactivity of IgG2 against rOMP40 in the second sampling of experimental calves was higher in herds with Charolaise and Limousine calves (p ≤ 0.001) and in crossbred calves (p ≤ 0.05). In the third sampling, serum IgG1 antibody reactivity against rOMP40 in Limousine calves was higher (p ≤ 0.05) in the experimental group. Among the other evaluated parameters, only SAA in the second sampling in the herd with Charolaise calves and heart rate in the herd with Limousine calves were significantly higher in the control calves (p ≤ 0.05). CONCLUSION: The application of HS to calves in all herds had an impact on specific reactivity in IgG1 and IgG2 classes against H. somni rOMP40 and rHsp60, antigens which were used for serum production.

Evaluation of the immunogenic properties of the recombinant Histophilus somni outer membrane protein 40 kDa (rOMP40).

BACKGROUND: Gram-negative bacterial infections are a serious problem in beef and dairy cattle. Bacterial outer membrane proteins (OMPs) play a pivotal role in cellular survival and the host-bacterium interaction. Histophilus somni OMP40 was identified as a porin with homology between its N-terminal amino acid sequence and the sequences of porins of other gram-negative bacteria The aim of this study was to produce recombinant H. somni OMP40 (rOMP40), optimize its production and evaluate its immunogenic properties in calves. The cross-reactivity of anti-rOMP40 antibodies were also checked. RESULTS: The highest overexpression of rOMP40 was demonstrated by Escherichia coli C41 using the autoinduction process. Double immunization of calves (20 µg rOMP40 per animal) induced a significant increase of anti-rOMP40 antibodies in the IgG1 (P ≤ 0.01) and IgG2 (P ≤ 0.01, after first immunization only) subclasses, but not IgM. ELISA revealed increased reactivity of the IgG against surface antigens of E. coli and Pasteurella multocida after the second immunization (P < 0.01). Cross reactivity of anti-rOMP40 antibodies with ~ 40 kDa antigens of most common gram-negative pathogens was shown by Western blotting. CONCLUSION: Immunization with H. somni rOMP40 induced a humoral response in cattle with broad cross-reactivity with similar antigens of other species of Pasteurellaceae and Enterobacteriaceae families and the delayed-type hypersensitivity reaction. The obtained results encourage further study to evaluate the protective effect of the produced protein as a subunit vaccine in cattle.

Staphylococcal Enterotoxin Genes in Coagulase-Negative Staphylococci-Stability, Expression, and Genomic Context.

In the current study, we screened a collection of coagulase-negative staphylococci (CoNS) isolates for orthologues of staphylococcal enterotoxins (SEs) involved in S. aureus-related staphylococcal food poisoning (SFP). The amplicons corresponding to SEs were detected in S. chromogenes, S. epidermidis, S. haemolyticus, S. borealis, S. pasteuri, S. saprophyticus, S. vitulinus, S. warneri, and S. xylosus. All amplicons were sequenced and identified as parts of known S. aureus or S. epidermidis SE genes. Quantitative real-time PCR allowed determining the relative copy number of each SE amplicon. A significant portion of the amplicons of the sea, seb, sec, and seh genes occurred at low copy numbers. Only the amplicons of the sec gene identified in three isolates of S. epidermidis displayed relative copy numbers comparable to sec in the reference enterotoxigenic S. aureus and S. epidermidis strains. Consecutive passages in microbiological media of selected CoNS isolates carrying low copy numbers of sea, seb, sec, and seh genes resulted in a decrease of gene copy number. S. epidermidis isolates harbored a high copy number of sec, which remained stable over the passages. We demonstrated that enterotoxin genes may occur at highly variable copy numbers in CoNS. However, we could identify enterotoxin genes only in whole-genome sequences of CoNS carrying them in a stable form at high copy numbers. Only those enterotoxins were expressed at the protein level. Our results indicate that PCR-based detection of enterotoxin genes in CoNS should always require an additional control, like analysis of their presence in the bacterial genome. We also demonstrate S. epidermidis as a CoNS species harboring SE genes in a stable form at a specific chromosome site and expressing them as a protein.

Changes of plasma fibronectin and fibronectin-fibrin complexes in dams of stillborn dairy calves.

BACKGROUND: Fibronectin (FN) is a large (450-500 kDa), multidomain and multifunctional glycoprotein existing in mammalian tissues. Some fibronectin (FN) molecular forms might be involved in biological processes occurring within the perinatal period, such as tissue remodeling, coagulation, and repair. RESULTS: In this study fibronectin (FN) and fibrinogen (Fb) concentrations and FN-fibrin complexes occurrence and its relative amounts with increasing high molecular masses were respectively determined by ELISA, heat precipitation, and SDS-agarose-immunoblotting methods. Plasma samples from three groups of dams with: 1) singleton stillborn calf without or with negligible autolytic changes in internal organs (DSBn), 2) singleton stillborn calf with advanced autolytic changes in internal organs (DSBa), 3) singleton live-born control calf (DC), and 4) a group of cows during mid to late lactation (LC) were analyzed. Maternal plasma FN concentration in the DSBn and DSBa groups was significantly lower than in the LC group. The plasma samples of DSBa showed a significantly lower FN concentration than in the DC group. Plasma Fb concentration was significantly higher in the DSBa and DSBn, than in the LC group. FN immunoblotting of the cow plasma samples revealed, besides an FN-dimer band, the presence of supramolecular FN-fibrin bands corresponding to FN-fibrin complexes with increasing molecular masses: up to 5 bands from 750 kDa to 1900 kDa in the DSBn and DSBa plasma samples, two bands of 750 and 1000 kDa in the DC group, and only the smallest one of 750 kDa in the LC group. CONCLUSIONS: The observed low FN concentration and occurrence of supramolecular FN-fibrin complexes (1000 kDa and more) in the maternal plasma comparing to cows in lactation might have been associated with periparturient changes in tissues. The presence in maternal plasma of high-molecular FN-fibrin complexes (1300-1900 kDa) arouse the question if this is the consequence of calf perinatal mortality.

Evaluation of the protective effect of immunization spf DBA/2J mice with selected bacterial, recombinant Hsp60 antigens during Salmonella Enteritidis challenge.

Salmonella Enteritidis is one of the most common causes of food poisoning in humans. Many attempts have been made to develop an effective vaccine against S. Enteritidis for use in poultry, but experiments aimed at the complete elimination of this pathogen from poultry farms have not provided satisfactory results. The development of new generation vaccines against salmonellosis, such as subunit vaccines based on heat shock proteins (HSPs), is strongly justified. The high immunogenicity of Hsp60 isolated from Procaryota, including Salmonella, has been suggested by the presence of IgG anti-Hsp60 antibodies in mice immunized with these proteins. The aim of the studies was to evaluate the protective effects of immunization with recombinant Hsp60 from selected gram-negative bacteria (S. Enteritidis, Escherichia coli, Pasteurella multocida, Histophilus somni) in spf DBA/2 J mice experimentally infected with S. Enteritidis. The study demonstrated that double subcutaneous immunization of mice with a dose of 10 µg rHsp60 induced a specific immune response of IgG antibodies in tested animals. The median lethal dose (LD50) for the murine model spf DBA/2 J orally infected with S. Enteritidis was estimated at 6.84 × 105 cfu/animal. Mice immunized with rHsp60 from gastrointestinal pathogens (S. Enteritidis and E. coli) showed better survival after experimental infection with a 3 × LD50 dose from S. Enteritidis, compared to animals immunized with proteins obtained from respiratory pathogens (P. multocida and H. somni). However, the log-rank analysis did not show significant differences in the survival rates between rHsp60-immunized mice and controls. S. Enteritidis was not isolated any less frequently from internal organs and faeces of rHsp60-immunized mice than from controls. Nevertheless, the level of haptoglobin (but not IL-6) was increased in all mice in which the presence of the pathogen was observed. Bacterial Hsp60 is an interesting candidate for a subunit vaccine, but its use in livestock animals must be further investigated.

Metabolomic studies as a tool for determining the post-mortem interval (PMI) in stillborn calves.

BACKGROUND: Perinatal mortality may vary between herds, but the cost of deaths are always higher than value of the calf. When diagnosing the cause of a calf's death it is important to determine when it occurred, before or after calving. Metabolomics is widely used to identify many human diseases, but quite rarely applied in veterinary science. The aim of this study was to compare the metabolic profiles of calves with different times of death and those of calves born alive. Into the study, twenty one healthy controls (singleton, normal assisted calving, born alive) and 75 stillborn (SB) calves (with a gestation length of ≥260 days, SB, or dead within 6 h of birth) were enrolled. Plasma and urine from SB and control calves were investigated by proton nuclear magnetic resonance based metabolomic methods. SB calves were divided into four PMI groups. One PMI group included calves that died after calving and the other groups - three comprised in utero deaths, based on pathophysiological changes (lung inflation, autolysis in internal organs, hemoglobin imbibition in the pleura and aortic arch). Partial Least Squares - Discriminant Analysis models based on plasma metabolites were calculated, reflecting assumed data clustering. RESULTS: Twenty six metabolites in plasma and 29 in urine changed significantly with PMI according to one way analysis of variance. Half the metabolites in plasma and the majority in urine increased with PMI. Six metabolites increased simultaneously in plasma and urine: acetate, sn-glycero-3-phosphocholine (GPC), leucine, valine, creatine, and alanine. CONCLUSIONS: Post-mortem changes in calves were associated with molecular variations in blood plasma and urine, showing the greatest differences for the group in which the post-mortem pathological changes were the most advanced. The results of the study show that evaluation of calf plasma or urine may be used as a diagnostic method for the determination of the PMI. Moreover, the metabolites, which unambiguously increased or decreased, can be used as potential biomarkers of PMI.

Selected prebiotics and synbiotics administered in ovo can modify innate immunity in chicken broilers.

BACKGROUND: A previous study showed that prebiotics and synbiotics administered in ovo into the egg air cell on the 12th day of incubation enhance the growth and development of chickens. However, the influence of this procedure on the development and efficiency of the innate immune system of broiler chickens is unclear. Therefore, the aim of this study was to evaluate whether the early (on the 12th day of embryo development) in ovo administration of selected prebiotics (inulin - Pre1 and Bi2tos - Pre2) and synbiotics (inulin + Lactococcus lactis subsp. lactis IBB SL1 - Syn1 and Bi2tos + L. lactis subsp. cremoris IBB SC1 - Syn2) influences the innate immune system. RESULTS: Chickens (broiler, Ross 308) that were treated with Pre1 exhibited a decreased H/L ratio on D7, but an increased H/L ratio was observed on D21 and D35. In the remaining experimental groups, an increase in the H/L ratio was observed on D21 and D35. The oxidative potential of leukocytes measured using the NBT test increased on D21 in Pre2 and Syn1 groups. The rate of the phagocytic ability of leukocytes increased in Pre1 and Syn1 groups on D21. The phagocytic index decreased in Pre1 and Syn2 groups on D21 and D35. Concurrently, the count of WBC in circulating blood decreased on D21 in Pre1, Pre2, and Syn1 groups. The hematocrit value was increased in Syn1 chickens on D21, in Pre1 chickens on D35, and in Syn2 chickens on both time points. CONCLUSIONS: Early in ovo treatment of chicken embryos with prebiotics and synbiotics may temporarily modulate not only the production/maturation of leukocytes but also their reactivity.

The evaluation of immunogenic impact of selected bacterial, recombinant Hsp60 antigens in DBA/2J mice.

Heat Shock Proteins (HSP) are highly conserved proteins that are widely spread throughout all organisms. They function in the cytoplasm as chaperones; however, they could be expressed on the cell surface. It has been shown that Hsp60 obtained from gram-negative bacteria are able to stimulate cells of the acquired and innate immune system. The aim of this study was the evaluation of the immunogenic properties of recombinant Hsp60 proteins derived from four common pathogenic bacteria: Escherichia coli, Histophilus somni, Pasteurella multocida and Salmonella Enteritidis. The analysis of the humoral immune response in DBA/2J mice hyperimmunized with selected rHsp60 revealed high levels of IgG rHsp60-antibody with the predominance of the IgG1 subclass, in the reaction with both homologous and heterologous antigens. The presence of IgG2a and IgG2b was also observed; however, no antibodies of subclass IgG3 were detected. The comparison of plasma IgG antibody reactivity of mice immunized with two different doses of rHsp60 (10/20 µg) showed that the lower dose was sufficient to induce a strong humoral response. The reactivity of the IgG rHsp60-antibody with whole bacterial cells showed a significantly higher reaction with H. somni compared with other pathogens. It was demonstrated that the addition of all rHsp60 with polymyxin B to the culture medium stimulated splenocytes isolated from hyperimmunized mice to release IL-1ß and IL-6. As a strong stimulator of the immune system, bacterial-origin Hsp60 seems to be an interesting potential component of subunit vaccines aimed at the development of protection for animals during infections caused by gram-negative bacteria.

Perinatal immuno/inflammatory responses in the presence or absence of bovine fetal infection.

BACKGROUND: It is known that the bovine fetus can mount an immune and inflammatory reaction to infection, but it is not known whether there is a contemporaneous maternal response. Nor is it known whether the response of calves which die perinatally, with or without infection, differs from that of live perinates. Hence, the objective of this study was to determine if acute phase reactant and immunoglobulin concentrations differed between calves (and their dams) in three groups: live calves (CC; n = 21) and dead calves with (PM INF+; n = 22) or without (PM INF-; n = 89) in utero infection. In calf plasma, serum amyloid A, haptoglobin, immunoglobulins M, G1 and G2 and interleukin-6 were measured. In dam serum, SAA and Hp was measured and in amniotic and abomasal fluid, IL-6 was measured. RESULTS: Live calves had higher plasma concentrations of SAA and IL-6 than dead calves with (PM INF+) or without (PM INF-) in utero infection. Calves in the PM INF-, but not PM INF+ group, had higher Hp concentrations than calves in the CC group. Calves in the PM INF+ group had higher IgG1 concentrations than calves in the PM INF- and CC groups. Except for higher IgG1 and IgG2 concentrations, biomarker values did not differ significantly between dead calves with or without in utero infection. Live calves had higher IL-6 concentrations in abomasal fluid compared to PM INF- calves. There were no significant differences in blood biomarker concentrations between dams of the three groups of calves. Amniotic fluid IL-6 concentrations were higher from the dams of control calves than the dams of uninfected calves. CONCLUSIONS: Differences in biomarkers (higher Hp and IgG1; lower SAA and IL-6) between perinatal mortalities and live perinates probably reflect differences between these two groups in age at sampling (SAA and IL-6) and in utero infection (IgG1). Out of the six analytes measured in calves, only IgG1 and IgG2 were biomarkers of (chronic) in utero infection.

Immune and inflammatory biomarkers in cases of bovine perinatal mortality with and without infection in utero.

The objective of this study was to compare acute-phase protein [serum amyloid A (SAA) and haptoglobin (Hp)] and immunoglobulin G1 and M concentrations in blood plasma of cases of bovine perinatal mortality due to infection in utero or traumotocia and in unexplained cases. Plasma samples were collected from 110 stillborn calves with bacterial infection (INF_B, n = 16), with viral or parasitic infection (INF_V/P, n = 31) during pregnancy, with lesions of fatal traumotocia (TRAUM, n = 22), and from unexplained deaths (UNEXPL, n = 41). Plasma immunoglobulin and SAA concentrations were measured by ELISA, and Hp concentrations were measured by the guaiacol method and ELISA. Concentrations of SAA in the INF_B group were higher than in the UNEXPL group and tended to be higher than in the INF_V/P group. A reference range (0-29 mg/L) was established for SAA in stillborn calves. Concentrations of Hp tended to be higher in the INF_B group compared with INF_V/P group. Concentrations of IgM tended to be higher in the INF_B group compared with the TRAUM and INF_V/P groups. Concentrations of IgG1 were numerically, but not significantly, higher in the INF_V/P and INF_B groups compared with the other groups. The results demonstrate upregulation of immune and inflammatory responses in stillborn calves exposed to bacterial infection in utero. The immune-inflammatory parameters did not differ between calves with viral or parasitic infections and traumotocia. These immune-inflammatory profiles did not contribute to the diagnosis of unexplained stillbirth. This is the first report of an elevated acute phase protein response in stillborn calves. Measurement of SAA and IgM concentrations may be used in the diagnosis of bacterial infections in stillborn calves.

Hydrolysis with Cucurbita ficifolia serine protease reduces antigenic response to bovine whey protein concentrate and αs-casein.

In the present study the effect of hydrolysis with non-commercial Cucurbita ficifolia serine protease on a reduction of the IgE and IgG binding capacity of whey protein concentrate and αs-casein was investigated. The intensity of the protein degradation was analyzed by the degree of hydrolysis, the free amino groups content and RP-HPLC. The ability to bind the antibodies by native proteins and their hydrolysates was determined using a competitive ELISA test. Deep hydrolysis contributed to a significant reduction of immunoreactive epitopes present in WPC. In the case of IgE and IgG present in the serum pool of children with CMA, the lowest binding capacity was detected in the 24 h WPC hydrolysate, where the inhibition of the reaction with native WPC was ≤23 and ≤60 %, respectively. The analysis of the IgG reactivity in the antiserum of the immunized goat showed that the lowest antibody binding capacity was exhibited also by 24 h WPC hydrolysate at a concentration of 1000 µg/ml where the inhibition of the reaction with nWPC was ≤47 %. One-hour hydrolysis of α-casein was sufficient to significant reduction of the protein antigenicity, while the longer time (5 h) of hydrolysis probably lead to the appearance of new epitopes reactive with polyclonal.

[Heat shock protein HSP60 and the perspective for future using as vaccine antigens]. / Bialka szoku cieplnego HSP60 i perspektywy ich wykorzystania jako antygenów szczepionkowych.

Heat Shock Proteins (HSPs) are widely spread in nature, highly conserved proteins, found in all prokaryotic and eukaryotic cells. HSPs have been classified in 10 families, one of them is the HSP60 family. HSP60 function in the cytoplasm as ATP-dependent molecular chaperones by assisting the folding of newly synthesised polypeptides and the assembly of multiprotein complexes. There is a large amount of evidence which demonstrate that HSP60 is expressed on the cell surface. Especially in bacteria the expression on the surface occurs constitutively and increases remarkably during host infection. HSP60 also play an important role in biofilm formation. In the extracellular environment, HSP60 alone or with self or microbial proteins can acts not only as a link between immune cells, but also as a coordinator of the immune system activity. This protein could influence the immune system in a different way because they act as an antigen, a carrier of other functional molecules or as a ligand for receptor. They are able to stimulate both cells of the acquired (naïve, effector, regulatory T lymphocyte, B lymphocyte) and the innate (macrophages, monocytes, dendritic cells) immune system. HSPs have been reported to be potent activators of the immune system and they are one of the immunodominant bacterial antigens they could be a good candidate for a subunit vaccine or as an adjuvant.

Changes in Skeletal Muscle Troponin T and Vitamin D Binding Protein (DBP) Concentrations in the Blood of Male Amateur Athletes Participating in a Marathon and 100 km Adventure Race.

This study assessed changes in creatine kinase (CK) activity and skeletal muscle troponin T (sTnT) concentrations in the blood, to estimate the degree of muscle degradation after exercise. In addition, the concentration of vitamin D binding protein (DBP) in the blood was assessed. DBP concentrations were measured in blood as a marker for plasma load by monomeric actin. The study included marathon (MR) participants and 100 km adventure race (AR) participants, who were examined before and after the race. There was a significant (16-fold) increase in CK activity among AR participants, and a significant increase in sTnT concentration-127% in the MR group and 113% in the AR group, while there was a statistically significant decrease in DBP concentration by 14% in the AR group. In addition, it was observed that the initial concentration of DBP in both groups was in a normal range, but was lower than the average population, and the DBP concentration in the AR group was lower than in the MR group. It was concluded that exhausting physical effort such as a marathon or adventure races causes muscle damage with a far stronger influence on sarcoplasm than on filaments. The short-term and slight reduction in the concentration of DBP in blood after such efforts may be due to the appearance of monomeric actin in plasma.

Production of staphylococcal enterotoxin R by Staphylococcus aureus strains.

Staphylococcal enterotoxin D and R (SED, SER) production was determined in 24 S. aureus strains harboring sed gene. Seven of them were not able to produce SED as evidenced by enzyme-linked immunosorbent assay and Western blotting. Sequencing revealed that all these strains harbor a variant of sed gene. Expression of SER was detectable in 22 out of 24 isolates, with variance in productivity ranging from ∼40 to 450 ng/mL. Out of the seven isolates not able to produce SED, three produced high amounts of SER (249-396 ng/mL), two produced less than 200 ng/mL of SER, and two were found to express no detectable amount of SER. Three of those were assigned to spa type t1677 with two being of agr type III and one of agr type I. One strain was t084, agr type II, one t603, agr type II, one 2920, agr type III, one t2920, agr type III, and one t5160, agr type I. Because conventional screening procedures involve only the detection of classical enterotoxins in food, the isolates not able to produce SED presented in this study could pose a threat to human health due to SER production.

Effects of Dominance and Sprint Interval Exercise on Testosterone and Cortisol Levels in Strength-, Endurance-, and Non-Training Men.

The aim of the study was to investigate the response of testosterone and cortisol to sprint interval exercises (SIEs) and to determine the role of dominance. The experiment was conducted in a group of 96 men, divided into endurance-training, strength-training, and non-training groups. Participants performed SIEs consisting of 5 × 10-s all-out bouts with a 50-s active recovery. Using the passive drool method, testosterone and cortisol concentrations were measured in saliva samples at rest at 10 min pre and 12 min post exercise. Participants' heart rate (HR) was measured during the whole exercise. Dominance was assessed by the participants before the study; the rating of perceived exertion (RPE) was measured immediately after each bout. The study showed that those who trained in endurance and strength sports had significantly lower mean HRs after five acute 10-s interval bouts than those in the non-training group (p = 0.006 and p = 0.041, respectively). Dominance has an inverse relation to changes in HR; however, it has no relation to hormone response. No significant differences were observed in testosterone and cortisol changes in the endurance-training, strength-training, and non-training groups after SIE (p > 0.05), which may indicate that the exercise volume was too low.

The relationship between the intensity of Gasterophilus intestinalis larvae infection and the serum and salivary humoral immune response in horses.

Infection with Gasterophilus intestinalis (botfly) larvae often occurs in horses. The aim of the study was to isolate the larvae of G. intestinalis and evaluate the serum and salivary humoral immune response using self-developed ELISA in G. intestinalis infected horses. Blood serum or saliva samples were taken from 125 infected horses and 54 uninfected slaughtered horses. The antigens from G. intestinalis larvae were used for development of ELISA in order to evaluate the intensity of G. intestinalis IgG, IgM, and IgA antibody reactivity in the serum or saliva of naturally infected horses and horses without larvae in the gastrointestinal tract (control group). Serum antibodies against second and third larvae's stadium antigens reacted significantly more intensively in infected than in healthy horses in IgG (p ≤ 0.001; p ≤ 0.05, respectively) and IgA (p ≤ 0.05;p ≤ 0.001, respectively) classes. Salivary IgG and IgA specific's antibody reactivity was significantly higher in horses with moderate (p ≤ 0.01) and severe infection (p ≤ 0.001) compared to the healthy horses. The determination of the G. intestinalis IgG and IgA antibody activity in saliva and serum may be used for detecting horses moderately and severely infested with larvae.

Role of Infection and Immunity in Bovine Perinatal Mortality: Part 2. Fetomaternal Response to Infection and Novel Diagnostic Perspectives.

Bovine perinatal mortality due to infection may result either from the direct effects of intrauterine infection and/or the fetal response to such infection, leading to the fetal inflammatory response syndrome (FIRS). Both intrauterine infection and FIRS, which causes multi-organ damage and involution of immune organs, compromise fetal survivability, sometimes fatally. Organ injury associated with FIRS may, in addition to causing fetal mortality, irreversibly compromise extrauterine adaptation of the neonate, a recognized problem in human fetuses. Diagnosis of intrauterine infection and of FIRS requires related, but independent analytical approaches. In addition to detection of pathogens, the immune and inflammatory responses of the bovine fetus may be utilized to diagnose intrauterine infection. This can be done by detection of specific changes in internal organs and the measurement of antibodies and/or elements of the acute phase reaction. Currently our ability to diagnose FIRS in bovine fetuses and neonates is limited to research studies. This review focuses on both the fetomaternal response to infection and diagnostic methods which rely on the response of the fetus to infection and inflammatory changes, as well other methods which may improve diagnosis of intrauterine infection in cases of bovine perinatal mortality.

Role of Infection and Immunity in Bovine Perinatal Mortality: Part 1. Causes and Current Diagnostic Approaches.

While non-infectious causes are more commonly diagnosed in cases of bovine perinatal mortality (PM), the proportion caused by infections is highly variable between studies (~5-35%); the reasons for this variation, and possible underestimation, are discussed. The most important pathogen-specific infectious causes of PM are bacteria (in particular, Bacillus licheniformis and Leptospira spp.), viruses (in particular BVDv) and a parasite (Neospora caninum). However, co-infection may occur in a small proportion of cases and in many cases no single pathogen is detected but gross or microscopic lesions of an inflammatory response are identified. Diagnosis is complicated by the criteria required to establish exposure, infection and causation. Additionally, pathogens can be classified as primary or secondary though such differentiation can be arbitrary. The majority of infectious cases of PM are due to in utero infections but postnatal infections (0-2 days) can also cause PM. Diagnosis of infectious PM is based on a systematic investigation of the herd health history and dam and cohort sampling and examination of the perinate and its placenta. Gross and histopathologic examinations and maternal/herd and perinate serology form the basis of current infectious PM investigations.

The Dynamics of Circulating Immune Complexes in Horses with Severe Equine Asthma.

Non-invasive diagnostic biomarkers of equine asthma syndrome (EAS) from blood or urine are sought. The aim of this study was to assess the absorbance of circulating immune complexes (CICs) during the exacerbation, remission, and treatment of an asthma episode and assess the potential usefulness of CIC levels in the diagnosis and monitoring of the disease. The control group, asthma group, and treated asthma group each contained six horses. Following an initial examination and group classification, the horses were kept in a dusty environment for seven days and then moved to an asthma-friendly environment for three weeks (the treated group received injections of glucocorticoids). Blood was collected at baseline and on the 1st, 2nd, 3rd, 7th, 14th and 30th days. CIC was measured using the modified Haskova method. The time points did not show significant statistical differences. There was a significant decrease in CIC in the treated group, and a significant increase in CIC in the non-treated group on day 30. CIC did not support the EAS diagnosis, although it may help in monitoring patients. To the best of the authors' knowledge, this is the first study to analyse the dynamics of CIC during environmental challenge, remission, and treatment.