Hepatic Coccidiosis in Wild Rabbits in Greece: Parasite Detection on Liver Imprints and the Associated Biochemical Profile.

(1) Background: Rabbit hepatic coccidiosis, caused by Eimeria stiedae, is a devastating disease with high morbidity and mortality rates. The disease is well described in rabbits, but little is known about E. stiedae infection in wild rabbits. In this study, we investigated the presence of E. stiedae infection in wild rabbits from the island of Lemnos, Greece, where this species is overpopulated, and the effects of infection on common hepatic biomarkers. (2) Methods: We used liver impression smears to detect the coccidian oocysts, and we defined the liver biochemical profile of the infected individuals. (3) Results: Overall, 13.3% of the liver imprints examined were positive for the presence of coccidial oocysts. The activities of liver enzymes, that is, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and glutamyltransferase (GGT), as well as globulins (GLOB), were increased while the concentrations of albumins (ALB), total proteins (TP) and the albumin to globulin (A/G) ratio were decreased in the infected individuals compared to the non-infected ones. (4) Conclusions: This study adds to the current knowledge on the pathogens affecting wild rabbits and those circulating in this population on the island of Lemnos, Greece. Moreover, we showed that E. stiedae infection exerts pathological effects on the hepatocyte integrity and liver function of wild rabbits, as reflected by the abnormal values of liver injury and dysfunction biomarkers.

Natural Co-Exposure to Borrelia burgdorferi s.l. and Anaplasma phagocytophilum: Unraveling the Hematological Profile in Sheep.

The occurrence of co-infected hosts and questing ticks with more than one tick-borne pathogen-as in the case of Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato-is expected in endemic regions. Their synergy-in terms of pathogenesis and disease severity-has been suggested previously in humans. Limited data exist on the clinicopathological alterations in co-infected sheep. In this study, we investigated the impact of A. phagocytophilum and B. burgdorferi s.l. seropositivity, alone and in combination, on the hematological parameters of naturally infected sheep. A complete blood count was performed, and indirect immunofluorescence assays were used to detect IgG antibodies against A. phagocytophilum and IgG and IgM antibodies against B. burgdorferi s.l. Single natural exposure to B. burgdorferi s.l. was characterized by low Packed Cell Volume (PCV) values and platelet (PLT) counts, while single exposure to A. phagocytophilum was characterized by low PCV values, low white blood cell (WBC) counts, and an increased risk for leukopenia and neutropenia. Co-exposure resulted in the most severe blood abnormalities; all the blood parameters decreased, and the sheep presented an increased risk for anemia. Our study showed that natural co-exposure to A. phagocytophilum and B. burgdorferi s.l. in sheep leads to more severe blood abnormalities and enhances the pathogenic processes. More studies are needed to clarify the possible background mechanisms.

Detection of Antibodies Against Toxoplasma gondii in Filter Paper-Dried Blood Dot Spots Compared with Serum in Pigs and Assessment of Variation Associated with Packed Cell Volume.

The aim of this study was to assess the agreement between anti-Toxoplasma gondii IgG antibody detection in serum and filter paper (FP) blood spots using the indirect immunofluorescence antibody assay (IFA) and to evaluate the potential impact of the packed cell volume (PCV) on antibody detection in FPs. A pair of a serum and an FP sample was collected from 96 sows at various farms in Greece, with previously identified high seropositivity and/or risk factors associated with high seropositivity against T. gondii. The PCV value was determined using the microhematocrit method. IFA was used for the detection of antibodies against T. gondii. T. gondii-specific antibodies were detected in 45.8% serum samples and 41.6% FP samples showing almost perfect agreement. Detection in FP samples presented high sensitivity (87.1-92.8%) and excellent specificity (100%) when compared with detection in serum, regardless of the PCV values. The findings of this study support the reliability of FPs for the evaluation of the serological status of swine against T. gondii. FPs could be a good alternative sample type compared with serum for large-scale epidemiological studies.

Effects of feeds, supplemented with humic substances and calcium carbonate, on performance, egg quality and heart rate variability in laying hens.

This study aimed to determine the effects of in-feed inclusion of humic substances (HS) and/or calcium carbonate (CaCO3) on the performance and welfare of laying hens. A total number of 144 Hy-Line Brown laying hens (55 weeks old) were randomly assigned to four treatments, T1-T4 (36 birds per treatment). T1 hens were fed on a control diet without HS or CaCO3, T2 hens were fed on the control diet + 2.00 g per bird per day CaCO3, T3 hens were fed on the control diet + 0.20% HS and T4 hens were fed on the control diet + 0.20% HS + 2.00 g/bird/day CaCO3. The experiment started after 15 days of adaptation and lasted 8 weeks. The parameters evaluated were percentage of hen-day egg production, food consumption, mortality, egg quality parameters and heart rate variability (welfare indicator). Hens in the T3 group showed a significantly lower feed intake than those in the other three groups, however, significantly higher daily egg production was recorded in groups T3 and T4 compared to T1 and T2. Eggshell quality characteristics were significantly improved by HS supplementation and eggs laid by hens in groups T1, and T4 presented paler yolk and shell color than those in the other groups. In conclusion, these results indicated that in-feed inclusion of HS had a beneficial effect on laying hens' productive performance including egg production and eggshells quality.

Prediction of ionized calcium concentration based on total calcium and protein levels in cattle and sheep.

Despite being important, there are no equations for prediction of ionized calcium (iCa) in sheep and cattle. The objectives of this study were i) to create equations for the calculation of serum iCa concentration based on the serum concentrations of total calcium (tCa), albumin (Alb) and total proteins (TP) and ii) to investigate whether predicted serum iCa values are beneficial in clinical practice. Serum samples from 30 sheep and 30 dairy cattle were used. Serum tCa was determined colorimetrically, while serum iCa was determined with an ion selective electrode method. Serum Alb and TP concentration were determined using bromo-cresol green and biuret methods, respectively. Ionized calcium was also calculated based on serum tCa, using regression analysis, and with two equations based on Alb and TP concentration. Bland-Altman plots were plotted to evaluate the agreement between measured and predicted iCa; Passing and Bablok (P - B) regression analysis was used to assess their agreement. The initial equations were corrected using the P - B generated equation and Bland - Altman plots were run to evaluate the level of agreement between measured and predicted iCa using the final equations. Six equations were finally created for cattle and 6 for sheep. The total bias exceeded 10.00% in all of them indicating that they are clinically unacceptable for iCa prediction especially when the predicted result is very close to the cut-off point of < 1.00 mmol L-1. So, it could be suggested that, when necessary, iCa concentration should be directly determined.

Surgical management of bilateral ear pinna lesions associated with traumatic aural hematoma in a 3-day-old goat kid.

Background: Aural hematomas are not uncommon in ruminants' clinical practice; however, there is a lack of information regarding their management in newborn ruminants, especially for complicated cases with rupture of the hematoma and secondary ear pinna necrosis. Case Description: A 3-day-old orphan goat kid was admitted due to swelling on the left ear pinna and trauma on the right pina caused by biting by other goats. The swelling on the left ear which was located at the convex surface was soft, painless, and fluid-filled, suggestive of aural hematoma located at the convex surface. The right pinna was swollen, bleeding, and extremely painful at palpation. The skin was necrotized at the distal 2/3rd of the convex surface and the 1/2 of the concave surface. Underneath the necrotized skin of the convex surface, blood, and blood clots were trapped, and there was a pocket between the remaining normal skin and the cartilage indicating possible rupture of aural hematoma. The kid was surgically treated under general anesthesia with xylazine and ketamine. The aural hematoma was drained by the convex surface using a Penrose tube after flushing the cavity with 2 mg dexamethasone. The trauma of the right pinna was left to heal by secondary intention after resection of all necrotized, edematous tissues, and blood clots. Post-surgery, the animal was treated with parenteral antibiotic administration and daily application of a topical antiseptic solution. The Penrose tube was removed after 5 days, and the animal recovered uneventfully. Conclusions: The present case indicates that aural hematoma can occur in newborn goat kids secondary to ear pinna biting and might evolve to pinna necrosis. In addition, tube drainage after flushing the cavity with corticosteroids appears to be an effective treatment approach without requiring bandaging post-operatively.

Neospora caninum and/or Toxoplasma gondii Seroprevalence: Vaccination against PCV2 and Muscle Enzyme Activity in Seropositive and Seronegative Pigs.

Neospora caninum and Toxoplasma gondii affect both humans and animals worldwide. To investigate their seroprevalence and differences in seropositivity between pigs vaccinated and unvaccinated against porcine circovirus 2 (PCV2), as well as differences in muscle enzyme activity between seropositive and seronegative pigs, blood samples were collected from 380 sows. Antibodies against T. gondii and N. caninum were detected by an indirect immunofluorescence antibody (IFA) assay, while the activities of creatine kinase (CK) and aspartate aminotransferase (AST) were biochemically assessed. Out of the 364 sows finally included in the study, 4.4%, 3.5%, and 0.5% were seropositive to T. gondii, N. caninum, or both. A significantly higher percentage of seropositivity against T. gondii and/or N. caninum in PCV2 unvaccinated pigs compared with vaccinated pigs was observed. Increased serum activities of CK and AST were detected in 71.43% and 100% of only against T. gondii (T+) and 63.64% and 90.91% of only against N. caninum (N+) seropositive sows, respectively, and were significantly higher compared to seronegative animals. T. gondii and N. caninum seropositivity, especially in presumed immunocompromised pigs, and the evidence of muscle damage highlight their importance as a zoonotic pathogen and animal model of human infection, respectively.

Potential Acute Renal Injury in Sheep with Bluetongue Serotype 4.

Bluetongue is a vector-borne disease with epidemic potential. Recently, outbreaks of Bluetongue were reported across Greece, caused by the Bluetongue virus (BTV) serotype 4. Regarding its pathogenesis, BTV infection involves various target organs with limited data referring to the kidneys. The objective of this study was to identify the possible impact of BTV infection on kidneys using common renal biomarkers. Urine and blood samples collected from 30 sheep with clinical signs of bluetongue (BTV sheep) and 30 clinically healthy sheep (normal sheep) from the same farms were finally selected and included in the study from an initial population of 47 sheep per group, based on the absence of active urine sediment. Complete urinalysis was performed and urine protein to creatinine ratio (UPC) and urine gamma-glutamyl transferase to creatinine (UGGTC) ratio were determined. Blood urea nitrogen (BUN), creatinine, total proteins, albumin (ALB), and inorganic phosphate (P) were determined in serum samples. UPC and UGGTC were significantly higher (p < 0.05) in BTV sheep compared to normal, whereas urine specific gravity (USG) was significantly lower (p < 0.05). Cylindruria was also detected in BTV sheep, and absence of azotemia in BTV and normal sheep. All these findings are indicative of renal tubular injury and/or dysfunction and suggestive of an association between BTV infection and acute damage of renal tissue.

Effects of housing systems on keel bone damage and egg quality of laying hens.

The aim of this study was to investigate whether keel bone damage is prevalent in laying hens in Greece. The study was conducted in three industrial farms using different housing systems: (a) enriched cages, (b) floor system, and (c) free-range system. One hundred hens per housing system, randomly selected, were evaluated for keel bone damages with the method of palpation. Complementarily, thirty eggs from each farm were selected for the measurement of egg weight, shape index, shell cleanness, shell color, shell breaking force, shell thickness, shell weight, egg yolk color, albumen height, and Haugh unit. The presence of keel bone damage was evident in all housing systems with the significantly highest occurrence being observed in the free-range system (50.00%), followed by enriched cages (24.00%) and floor system (7.00%). Eggs from all three systems had significant differences in all estimated egg quality parameters apart from shell color and Haugh unit.

Reference values of Schirmer tear test in sheep and the effect of season on the test results.

The aim of this study was to determine the reference range of Schirmer tear test (STT) values in sheep using Greek indigenous and mixed breeds and to investigate the potential effect of breed and season on these values. Ninety (30 Chios, 30 Florina and 30 Lacaune mixed breed) ewes aged 2 years or more were used for the study. The aqueous portion of the tear film was measured using Schirmer tear test (STT) commercial strips bilaterally by the same investigator with the animal in standing position. The average STT value recorded was 18.45 ± 3.93 mm/min and the range 10.8-26.2 mm/min. STT was significantly affected by the season and the recorded values were significantly higher in summer compared to winter. The breed had no significant effect on tear secretion. The results of the present study provide a reference range of STT values in sheep and indicate that tear secretion is significantly affected by the season.