Successful Transmammary Treatment of Babesia gibsoni Infection in Newborn Puppies after the Administration of Malarone®, Azithromycin, and Artesunate to a Lactating Dam.

Babesia gibsoni is a parasitic protozoan transmitted through tick bites and can cause severe disease in dogs. It can also be transmitted through direct contact with infected blood during dog fights, blood transfusions, and from dam to offspring during the perinatal period, resulting in stillborn or dead newborn puppies. This study aimed to determine the incidence of infection, the viability of newborn puppies, and the degree of B. gibsoni transmission from infected dam to offspring during pregnancy and lactation. Using PCR-based molecular methods, B. gibsoni infection in a pregnant American Pit Bull Terrier and her newborn puppies was confirmed. The incidence of B. gibsoni infection in the litter reached 75%. Out of eight puppies, six were infected with B. gibsoni, and one died. A therapeutic protocol comprising Malarone®, azithromycin, and artesunate was administered to a lactating B. gibsoni-positive bitch. By day 77 after birth, three out of five positive puppies showed negative PCR tests for B. gibsoni, indicating successful treatment through breast milk during nursing. In the two remaining positive puppies, therapy was started and parasitemia was successfully eliminated.

The Effect of Enterococcus faecium AL41 on the Acute Phase Proteins and Selected Mucosal Immune Molecules in Broiler Chickens.

Probiotic bacteria, including the Enterococcus faecium strain, can improve intestinal mucosal health by several mechanisms, including modulation of the immune response, as well as by improving the protective function of the epithelial barrier. In this study, we tested the effect of Enterococcus faecium AL41 on the acute phase proteins response (blood), gene expression of selected molecules of mucosal immunity (immunoglobulin A, mucin-2, insulin-like growth factor 2) and mucus production (all parts of the small intestine) in broilers. Eighty broiler chicks were divided into two groups: a control and E. faecium AL41 (birds were inoculated with AL41 for 7 days) group. The whole experiment lasted 11 days. Our results revealed that the administration of E. faecium AL41 had no substantial effect on the concentrations of acute phase proteins, but we recorded a significant increase in ß- and γ-globulin fractions at the end of the experiment, which may indicate an improvement in the immune status. A significant prolonged stimulatory effect of E. faecium AL41 on the relative expression of molecules (immunoglobulin A, mucin-2) as well as on the dynamic of mucus production in the chicken intestine was observed. In addition, AL41 significantly reduced the total number of enterococci in the cecum and faeces.

Clinical Efficacy and Safety of Malarone®, Azithromycin and Artesunate Combination for Treatment of Babesia gibsoni in Naturally Infected Dogs.

Babesia gibsoni is a tick-borne protozoal blood parasite that may cause hemolytic anemia, thrombocytopenia, lethargy, and/or splenomegaly in dogs. Many drugs have been used in management of canine babesiosis such as monotherapy or combined treatment, including diminazene aceturate, imidocarb dipropionate, atovaquone, and antibiotics. This report examines the effectiveness and safety of Malarone®, azithromycin (AZM) and artesunate (ART) combination for the treatment of babesiosis in dogs naturally infected with Babesia gibsoni. Twelve American Pit Bull Terriers were included in the experiment. Examined dogs underwent clinical and laboratory analysis including hematology and biochemistry profile and serum protein electrophoresis. After diagnosis, the dogs received combined therapy with Malarone® (13.5 mg/kg PO q24 h), azithromycin (10 mg/kg PO q24 h) and artesunate (12.5 mg/kg PO q24 h) for 10 days. The combined treatment improved hematology and biochemical parameters to the reference range gradually during the first 14 days already, resulting in the stable values until day 56 after treatment. No clinically apparent adverse effects were reported during treatment and monitoring. No relapses of parasitemia were detected in control days 180, 360, 540 and 720 in all dogs. Results of the study indicate that the combined treatment leads to successful elimination of parasitemia in chronically infected dogs with B. gibsoni.

Changes in the Serum Protein Fractions in Goats after Treatment of Natural Gastrointestinal Parasite Infections.

Gastrointestinal parasitic infections in small ruminants belong to major health problems. The regulation of gastrointestinal infections in goats and the responses developed against them appear to be different from those observed in sheep. In the literature, there is a lack of data on the effect of gastrointestinal parasitic infections on the serum protein profile in goats. Therefore, the aim of the study was to determine the electrophoretic pattern of serum proteins in goats naturally infected with gastrointestinal parasites and to compare the changes in the total serum proteins and serum protein fractions (albumin and α 1-, α 2-, ß-, and γ-globulins) obtained after antihelminthic treatment. Eight adult female goats of the white shorthaired breed from a small dairy goat farm at the age ranging between 3 and 5 years with average body weight 35.4 ± 3.2 kg and body condition score (BCS) from 1.5 to 2.5 were used in the study. The serum proteins in goats were separated into five fractions: albumin, α 1- and α 2-globulins, and ß- and γ-globulins. Significant changes after treatment were found in the relative concentrations of albumin (P < 0.01) and α 2- (P < 0.05), ß- (P < 0.001), and γ-globulins (P < 0.01), as well as albumin/globulin ratio (P < 0.01). The mean concentration of total serum proteins was, after the antiparasitic treatment, significantly higher. Among the globulin fractions, the γ-globulin fraction contributed most significantly to these changes (P < 0.001). The results presented in the study suggest a significant effect of antiparasitic treatment in goats on the synthesis of blood serum proteins and on the changes of the proportion of serum protein fractions.

Serum protein electrophoretic pattern in piglets during the early postnatal period.

The pattern of serum proteins, the typical features of the electrophoretogram in newborn piglets and during their postnatal development is not completely described. Therefore, the aim of this study was to characterize the changes in serum protein electrophoretic pattern and features of the electrophoretograms during the early postnatal period. Significant changes during the monitored period were found in all evaluated parameters (P < 0.001). The most marked changes were observed mainly in the period before weaning. The concentrations of total proteins, albumin and γ-globulins were before colostrum intake low, γ-globulins represented the smallest proportion of protein fractions. The proportion of α1-globulins was after birth a dominant protein fraction. Significant increase of total proteins, α2-, ß- and γ-globulins and decrease of α1-globulins was found 2 days after colostrum intake. The albumin and A/G values increased after birth gradually until weaning. After weaning a significant changes were found in absolute concentrations of total protein and albumin, and in relative values of ß-globulin fractions. Presented results showed marked developmental alterations in the serum protein pattern in piglets along with the age. The study also brings new knowledge in the field of description of typical features of electrophoretograms in the observed period of piglet's life.

Differences in serum protein electrophoretic pattern in dogs naturally infected with Babesia gibsoni and Babesia canis.

Canine babesiosis may cause several hematological and biochemical changes, but only limited studies are available regarding the possible differences of changes in animals infected by different Babesia parasites. The study focused on the evaluation of the differences in serum protein electrophoretic pattern between dogs naturally infected with B. gibsoni (17 dogs) and B. canis (40 dogs). The mean values of total proteins, ß1-, ß2- and γ-globulins were in dogs infected with B. gibsoni significantly higher (P < 0.05 and P < 0.001) than in dogs infected with B. canis. The relative concentrations of albumin, α1-, α2-globulins and the A/G ratios were in the B. gibsoni infected dogs significantly lower (P < 0.001), no significant differences were found in the relative concentrations of ß1- and ß2-globulins. Significant differences were found in most of the evaluated parameters when comparing the results in relation to the form of B. canis infection to B. gibsoni infection. Hematological indices showed significant differences between dogs infected with B. gibsoni and the complicated form of B. canis infection. In conclusion, the obtained results suggest differences in the changes of serum protein electrophoretic pattern between dogs infected with both Babesia species and thus, in the response to the infection caused by various Babesia parasites.

Clinical and Diagnostic Significance of Lactate Dehydrogenase and Its Isoenzymes in Animals.

Lactate dehydrogenase (LDH) is widely distributed enzyme in cells of various living systems where it is involved in carbohydrate metabolism catalyzing interconversion of lactate and pyruvate with NAD+/NADH coenzyme system. Cells of tissues are direct source of lactate dehydrogenase isoenzymes that are naturally distributed in blood plasma/serum of animals and humans producing characteristic profile. This profile depends on intracellular isoenzyme concentration in all tissues that contribute to the common pool of lactate dehydrogenases in plasma/serum as a consequence of natural cell degradation. LDH is widely distributed in the body, high activities are found in the heart, liver, skeletal muscle, kidney, and erytrocytes, whereas lesser amounts are found in the lung, smooth muscle, and brain. Because of its widespread activities in numerous body tissues, LDH is elevated in a variety of disorders. There are many conditions that contribute to increased activity of LDH. An elevated total LDH value is a rather nonspecific finding. Therefore, LDH assays assume a more clinical significance when separated into isoenzyme fractions. The activity of LDH and its serum and tissue patterns and composition show great variations between the species. These differences do not allow using catalytic activities of LDH isoenzymes from one species to another. Instead, the pattern of serum LDH isoenzymes should be interpreted in respect to its species origin that is important in particular in veterinary medicine. Determination of total LDH activity and its isoenzyme pattern in serum of mammals had become one of the biochemical indicators in the assessment of organ disorders. When the content of cells is released from tissue to plasma, as on cell injury, the LDH isoenzyme pattern of the serum changes in favour of the profile of the affected organ (tissue) that can be used in the diagnostic practice.

Changes in the Acute-Phase Protein Concentrations and Activities of Some Enzymes in Pigs Following the Repair of Experimentally Induced Articular Cartilage Defects Using Two Types of Biocement Powder.

The objective of the study was to assess the usefulness of acute-phase proteins (APPs) and serum enzymes in the evaluation of post-operative state after cartilage reconstruction in an animal model (Sus scrofa domesticus). Fifteen clinically healthy female pigs were evaluated during the first 30 days after the repair of experimentally induced articular cartilage defects using two types of biocement powders. Animals were divided into groups according to the type of biocement powder used: CAK-with amino acids (n = 6), C-without amino acids (n = 6) and the control group (Ctr) was without biocement (n = 3). The concentrations of selected APPs-serum amyloid A (SAA), haptoglobin (Hp) and C-reactive protein (CRP), and the activities of some serum enzymes-creatine kinase (CK), alkaline phosphatase (AP), and lactate dehydrogenase (LD) were measured one day before the surgery and on days 7, 14, and 30 after the surgical intervention. The most significant changes during the evaluated period were observed in the concentrations of SAA (p < 0.001) and Hp (p < 0.001), with marked increase of values 7 days after surgery. There was a numerical, but not statistically significant, difference between CAK, C and Ctr groups (p > 0.05). Marked variations were observed also in the activities of the evaluated enzymes, with the most significant changes in the activity of AP in the CAK group (p < 0.001). Presented results suggest possible usefulness of some APPs and serum enzymes in the evaluation of post-operative inflammatory state after the reconstruction of articular cartilage defects.

Changes of total protein and protein fractions in broiler chickens during the fattening period.

BACKGROUND AND AIM: Blood proteins in birds serve as an important indicator in the evaluation of health status and represent a basis in general biochemistry allowing the identification of metabolic alterations. The objective of this study was to evaluate the protein profile in broiler chickens extended by the concentrations of serum protein fractions at different periods of fattening. MATERIALS AND METHODS: Into the evaluation, we included 24 clinically healthy Ross 308 line meat-type chickens at the age of 2 days. Blood samples were taken on day 4, 18, 32, and 46 of fattening always from six randomly selected chickens. Chickens were fed with a commercial starter, grower, and finisher feeds. The concentrations of total serum protein and protein fractions were evaluated. RESULTS: Various significant changes in the proportion of the individual protein fractions were found during the observed period except for the beta-globulins in all protein fractions and the albumin/globulin (A/G) ratio. At the beginning of the fattening period, the relative concentrations of albumin, α1-globulins, and A/G ratio were significantly lower and the values of α2- and γ-globulins significantly higher (p<0.05). The values of pre-albumin fraction were found as a small band preceding the albumin fraction differed significantly between the different age groups of chickens (p<0.05). The total serum protein concentrations showed higher values in older broilers; the significantly highest mean value was recorded on day 32 of fattening. CONCLUSION: The results suggest that fattening and age of broilers influences not only the production patterns, metabolic processes, and lipid and mineral profile but also the parameters of protein profile. However, seeing that some contradictory data exist regarding the number and size of globulin fractions in chickens, further analyses are needed.

The Serum Protein Profile and Acute Phase Proteins in the Postoperative Period in Sheep after Induced Articular Cartilage Defect.

Although several new implants have been developed using animal studies for the treatment of osteochondral and cartilage defects, there is a lack of information on the possible metabolic and biochemical reactions of the body to the implantation of biomaterials and cartilage reconstruction. Therefore, this study was aimed at evaluating the serum protein pattern and the alterations in the concentrations of selected acute phase proteins in five clinically healthy female sheep before and after the reconstruction of experimentally induced articular cartilage defects using polyhydroxybutyrate/chitosan based biopolymer material. The concentrations of total serum proteins (TSP), protein fractions, and selected acute phase proteins-serum amyloid A (SAA), haptoglobin (Hp), and C-reactive protein (CRP)-were measured before and on days seven, 14, and 30 after the surgical intervention. The TSP concentrations showed no marked differences during the evaluated period. Albumin values decreased on day seven and day 14 after surgery. In the concentrations of α1-, α2-, ß-, and γ2-globulins, a gradual significant increase was observed during the postoperative period (p < 0.05). The γ1-globulins decreased slightly seven days after surgery. The concentrations of SAA, Hp, and CRP increased significantly after the surgical intervention with a subsequent decrease on day 30. Presented results suggest marked alterations in the serum protein pattern after surgical intervention.

Stability of ovine serum protein fractions under different storage conditions.

We evaluated the effect of different storage conditions on the concentration of serum total proteins and their electrophoretic fractions in sheep. Blood samples were collected from 12 female Comisana sheep, and each serum obtained was divided into 4 aliquots. The first aliquot was analyzed immediately after separation (T0); the second was refrigerated at 4°C and analyzed after 24 h (T1), 48 h (T2), and 72 h (T3); the third was stored at -20°C immediately after centrifugation and analyzed after 2 days (T4); and the last aliquot was stored at -20°C after 24-h refrigeration and analyzed after 1 wk (T5). The concentrations of serum total proteins and their electrophoretic fractions (albumin, α-, ß1-, ß2-, and γ-globulins) was determined. A statistically significant ( p < 0.05) effect of storage conditions on all serum electrophoretic protein fractions and on the albumin-to-globulin (A/G) ratio was found. The values of albumin, α-globulins, and A/G ratio increased at T4 with respect to T0. The levels of α-globulins were higher at T1 than T0; the ß1-globulins levels changed at T3, whereas the ß2- and γ-globulins values as well as the A/G ratio changed at T2 and T3. At T4, we found an increase in the serum concentrations of albumin, α1-globulins, and on the A/G ratio, and a decrease of γ fraction. At T5, the concentration of α-globulins was higher than at T0. Hence, we recommend electrophoretic analysis of fresh ovine serum samples or of samples refrigerated for no more than 24 h to obtain accurate data.

The effect of storage temperature and time on the concentrations of bovine serum amyloid a and its mammary associated isoform.

The objective of this study was to evaluate the effect of storage under various conditions on the concentrations of major bovine acute phase protein-serum amyloid A, and its mammary isoform. Blood samples were taken from seven clinically healthy calves, and milk samples from six clinically healthy dairy cows. The harvested blood serum and the milk samples were fractioned into aliquots. One aliquot was analyzed on the day of collection without storage. The second aliquots were stored at 4°C for 1 day, the remaining aliquots were kept frozen at -18°C for 2, 7, 14, and 21 days, and then analyzed. Blood serum was analyzed for serum amyloid A (SAA). The concentrations of mammary isoform of SAA (M-SAA) were measured in milk samples. Over time, the concentrations of SAA in serum showed a tendency of significant decrease during storage at -18°C (P < 0.01). Similarly, the values of M-SAA decreased significantly in samples maintained at freezer temperatures (P < 0.001). In the refrigerated samples, we found non-significantly lower values of SAA, as well as M-SAA. Presented results indicate that the storage temperature and duration may markedly affect the concentrations of bovine SAA and M-SAA.

The effect of chronic respiratory diseases on acute phase proteins and selected blood parameters of protein metabolism in calves.

This work was aimed at evaluation of the influence of chronic respiratory diseases on selected acute phase proteins (APPs), and on the concentrations of some parameters of protein metabolism in calves, as well as at evaluation of APPs as possible prognostic indicators in these diseases. We compared the results from 27 sick calves with results from 15 clinically healthy calves. Blood samples were analysed for concentrations of haptoglobin (Hp), serum amyloid A (SAA), total proteins, albumin, urea, creatinine, and total immunoglobulins. In calves with chronic respiratory diseases we found significantly higher concentrations of Hp and SAA than in healthy animals (P < 0.001 and P < 0.01, respectively). Moreover, in sick calves, concentrations of these parameters were significantly higher in died or euthanised calves compared with calves in improved health status during therapy (P < 0.05 and P < 0.01, respectively). The concentrations of Hp in evaluated calves correlated significantly with the values of SAA (P < 0.001). In sick animals we found also significantly higher mean concentrations of total proteins and total immunoglobulins (P < 0.001 and P < 0.001, respectively), and significantly lower mean concentrations of albumin and creatinine (P < 0.01 and P < 0.01, respectively). Our findings show that in cases of chronic respiratory diseases, there is also an increased production of the mentioned APPs, and their evaluation could be useful in the determination of prognosis in sick calves. Moreover, the results suggest that these diseases could also significantly affect the concentrations of some other biochemical parameters. Thus, these results could be an important contribution to the possible laboratory diagnosis of respiratory problems in cattle.