Neospora caninum in Aborted Bovine Fetuses in Trakya Region, Turkey - Histopathological, Immunohistochemical and Molecular Detection

Background: Being the major cause of bovine abortion in the world, Neosporosis is considered to be a very important protozoal infection in dairy cattle. Vertical transplacental transmission is the major route of the infection causing either abortion or birth of calves with persistent infection. As the seropositivity in individual cows and in fetal serology only indicate exposure to the protozoa, the diagnosis of the infection has to be based on histopathology of aborted fetuses. Additional techniques such as immunohistochemistry (IHC) and PCR are required for the detection of the etiological agent. The purpose of the current study was to diagnose Neospora caninum infection in aborted bovine fetuses in Trakya Region of Turkey. For this purpose, serological, histopathological, IHC, and PCR methods were used. Materials, Methods & Results: The blood samples and the fetuses of 55 aborted dairy cattle from various farms located in 3 provinces of Trakya, Turkey constituted the material of the present study. The sera obtained from the blood samples were tested using a Neospora caninum Antibody Test Kit cELISA and anti-N. caninum antibodies were detected in the sera of the dams of the 8 aborted fetuses (8/55; 14.54%). Following the necropsy, samples from the brain, heart, liver, lung, kidney, spleen, and placenta of 55 fetuses were routinely processed for histopathological examination and evaluated under a light microscope. Nonsuppurative encephalitis (15/55; 27.27%), necrosis (5/55; 9%) and gliosis (1/55; 1.8%) in the brain, mild to severe nonsuppurative myocarditis and epicarditis (14/55; 25.45%), and portal to mid-zonal nonsuppurative hepatitis (13/55; 23.63%) were the relevant findings. PCR analysis was performed on fresh frozen fetal tissues. Nested PCR detected N. caninum DNA in the brain, heart, liver, lung, and kidney tissues of 6 fetuses (6/55; 10.9%). IHC was performed on the brain, heart, and liver tissues of all the fetuses using avidin-biotin-complex peroxidase method. Immunoreactivity was observed in the brain of 1 fetus (1/55; 1.8%). Discussion: In the present study, histopathological, immunohistochemical and PCR analyses were performed to detect N. caninum in 55 spontenously aborted bovine fetuses in Trakya Region, Turkey. Histopathologic hallmark of the study was nonsuppurative inflammation found mostly in the brain, heart and liver followed by kidneys and lungs. No protozoa was observed in the microscopic examination supporting the fact that definitive diagnosis of N. caninum infection requires ancillary techniques such as IHC and PCR. Nested PCR detected N. caninum DNA in the tissues of 6 fetuses (6/55; 10.9%). Brain was the most reliable organ for detection by PCR (6/6; 100%), compatible with the previous reports. IHC diagnosis revealed only 1.8% positivity in the present study which was remarkably lower than found in the previous studies. Even though histopathology in conjunction with IHC are accepted as the “gold standard” methods to detect N. caninum infection in aborted bovine fetuses, there are studies claiming that IHC is relatively insensitive in the diagnosis of neosporosis as parasite numbers can be low and thus, false negative results can be obtained. Other factors affecting the sensitivity of the technique are thoroughly discussed by many authors. Supportively, the findings of the current study showed that using both IHC and PCR as complementary techniques, increases the success of detection of N. caninum as recommended in previous studies. In conclusion, the present study demonstrated the first molecular diagnosis of Neospora caninum infection in bovine aborted fetuses in Trakya Region of Turkey which has a critical geographical location bordering Europe. Keywords: bovine abortion, protozoal infection, histopathology, immunohistochemistry, PCR, Neospora caninum

Effects of oxygen supplementation during the last stage of incubation on broiler performance, ascites susceptibility and some physiological traits

The present study was conducted to determine the possible use of supplemental oxygen treatment during the last 3 days of incubation on broiler performance, ascites susceptibility and some physiological traits. On the 18th day of incubation, fertile eggs were divided into two groups. From 18 to 21 d, the eggs were placed in two hatcher cabinets: one being a control at normal atmospheric conditions, $20.9 \pm 0.5$% at $754 \pm 2$ mm Hg and the other supplemented with oxygen at $23.0 \pm 0.5$%. Oxygen supplementation had a significant effect on initial chick weight. At 6 weeks of age, body weight, growth rate, and feed consumption increased as partial pressure of O$_{2}$ rose from 20.9 to 23.0%. Feed conversion ratio decreased with the increasing partial pressure of oxygen. In the present study, general mortality and mortality due to ascites did not differ between the groups. At 5 weeks of age, oxygen supplementation did not cause significant changes in RBC (red blood cell count), Hb (hemoglobin) and Glucose. However, PCV (hematocrit) increased significantly as the partial pressure O$_{2 }$ rose from 20.9 to 23.0%. There were no differences in the right ventricle, left ventricle + septum, total ventricle and RV:TV (right ventricle:total ventricle) ratio between the oxygen supplemented and control group.Effets d'un apport supplémentaire d'oxygène durant le dernier stade d'incubation sur les performances du poulet de chair, la formation d'ascite et quelques caractères physiologiques. La présente étude a été conduite pour évaluer l'utilisation possible d'un apport supplémentaire d'oxygène pendant les trois derniers jours d'incubation sur les performances du poulet de chair, l'apparition d'ascite et quelques paramètres physiologiques. Au 18$^{\rm e}$ jour d'incubation, les œufs fertiles ont été divisés en deux groupes. De 18 à 21 jours, les œufs ont été placés dans deux éclosoirs, l'un (groupe témoin) sous conditions atmosphériques normales ($20,9 \pm 0,5$ % O$_{2}$ à $754 \pm 2$ mmHg) et l'autre (groupe expérimental) sous atmosphère enrichie en oxygène à $23,0 \pm 0,5$ %. L'apport d'oxygène a eu un effet significatif sur le poids initial des poussins. A six semaines d'âge, le poids corporel, la vitesse de croissance et la consommation d'aliments ont été accrus avec la pression partielle en oxygène la plus élevée, alors que l'indice de consommation a été diminué. La mortalité en général et la mortalité due à l'ascite en particulier n'ont pas différé entre les groupes. A cinq semaines d'âge, l'apport d'oxygène n'a provoqué aucun changement significatif pour la numération des globules rouges, le taux d'hémoglobine et de glucose. Toutefois, l'hématocrite a été sensiblement augmenté lorsque la pression partielle d'O$_{2}$ est passée de 20,9 à 23,0 %. Aucune différence significative n'a été observée entre les deux groupes pour le poids du ventricule droit, le poids du ventricule gauche + septum interventriculaire, le poids ventriculaire total et le rapport du poids du ventricule droit au poids ventriculaire total (VD/VT).

Assessment of neutrophil to lymphocyte ratio and mean platelet volume in pediatric familial Mediterranean fever patients

Background: Blood neutrophil to lymphocyte ratio (NLR) and mean platelet volume (MPV) both have been used as a simple marker of inflammation in many disorders. Here, we aimed to investigate the relationship between NLR, MPV, and familial Mediterranean fever (FMF). Materials and Methods: In this retrospective study, the files of FMF patients in pediatric rheumatology outpatient clinic were reviewed. There were 160 participants (68.4%) in the FMF patient group and 74 participants (31.6%) in the control group. Ninety of patients were in attack-free period, and 70 were in attack period. Results: The highest values of NLR were found in the patients at attack period. Patients in attack-free period and the participants in control group had similar levels of NLR (1.71 ± 0.83 and 1.91 ± 1.86 respectively) (P = 0.457), and they had lower ratios than the patients did at attack period (4.10 ± 3.11) (P < 0.001 for both). There was no significant difference between MPV values of attack patients (8.35 ± 4.91) and attack-free patients (8.43 ± 1.15) (P = 0.074). MPV values of attack patients and attack-free patients were significantly higher than control group (7.99 ± 0.81) (P < 0.001 for both). Conclusion: NLR ratio may indicate FMF attack period. Since there was no significant difference between attack-free patients and control groups, NLR ratio cannot be used as a subclinical inflammation marker. However, NLR could be a useful predictor of inflammation in FMF patients. On the other hand, since our attack and attack-free patients have similar MPV values and both had greater MPV values than control group, we suggest that MPV may be used to show subclinical inflammation

Neospora caninum in Aborted Bovine Fetuses in Trakya Region, Turkey - Histopathological, Immunohistochemical and Molecular Detection

Background: Being the major cause of bovine abortion in the world, Neosporosis is considered to be a very important protozoal infection in dairy cattle. Vertical transplacental transmission is the major route of the infection causing either abortion or birth of calves with persistent infection. As the seropositivity in individual cows and in fetal serology only indicate exposure to the protozoa, the diagnosis of the infection has to be based on histopathology of aborted fetuses. Additional techniques such as immunohistochemistry (IHC) and PCR are required for the detection of the etiological agent. The purpose of the current study was to diagnose Neospora caninum infection in aborted bovine fetuses in Trakya Region of Turkey. For this purpose, serological, histopathological, IHC, and PCR methods were used. Materials, Methods &amp; Results: The blood samples and the fetuses of 55 aborted dairy cattle from various farms located in 3 provinces of Trakya, Turkey constituted the material of the present study. The sera obtained from the blood samples were tested using a Neospora caninum Antibody Test Kit cELISA and anti-N. caninum antibodies were detected in the sera of the dams of the 8 aborted fetuses (8/55; 14.54%). Following the necropsy, samples from the brain, heart, liver, lung, kidney, spleen, and placenta of 55 fetuses were routinely processed for histopathological examination and evaluated under a light microscope. Nonsuppurative encephalitis (15/55; 27.27%), necrosis (5/55; 9%) and gliosis (1/55; 1.8%) in the brain, mild to severe nonsuppurative myocarditis and epicarditis (14/55; 25.45%), and portal to mid-zonal nonsuppurative hepatitis (13/55; 23.63%) were the relevant findings. PCR analysis was performed on fresh frozen fetal tissues. Nested PCR detected N. caninum DNA in the brain, heart, liver, lung, and kidney tissues of 6 fetuses (6/55; 10.9%). IHC was performed on the brain, heart, and liver tissues of all the fetuses using avidin-biotin-complex peroxidase method. Immunoreactivity was observed in the brain of 1 fetus (1/55; 1.8%). Discussion: In the present study, histopathological, immunohistochemical and PCR analyses were performed to detect N. caninum in 55 spontenously aborted bovine fetuses in Trakya Region, Turkey. Histopathologic hallmark of the study was nonsuppurative inflammation found mostly in the brain, heart and liver followed by kidneys and lungs. No protozoa was observed in the microscopic examination supporting the fact that definitive diagnosis of N. caninum infection requires ancillary techniques such as IHC and PCR. Nested PCR detected N. caninum DNA in the tissues of 6 fetuses (6/55; 10.9%). Brain was the most reliable organ for detection by PCR (6/6; 100%), compatible with the previous reports. IHC diagnosis revealed only 1.8% positivity in the present study which was remarkably lower than found in the previous studies. Even though histopathology in conjunction with IHC are accepted as the “gold standard” methods to detect N. caninum infection in aborted bovine fetuses, there are studies claiming that IHC is relatively insensitive in the diagnosis of neosporosis as parasite numbers can be low and thus, false negative results can be obtained. Other factors affecting the sensitivity of the technique are thoroughly discussed by many authors. Supportively, the findings of the current study showed that using both IHC and PCR as complementary techniques, increases the success of detection of N. caninum as recommended in previous studies. In conclusion, the present study demonstrated the first molecular diagnosis of Neospora caninum infection in bovine aborted fetuses in Trakya Region of Turkey which has a critical geographical location bordering Europe. Keywords: bovine abortion, protozoal infection, histopathology, immunohistochemistry, PCR, Neospora caninum

A Reconfigurable Nested Ring-Split Ring Transmitarray Unit Cell by Microfluidic Technology

This paper presents a reconfigurable circularly polarized transmitarray unit cell by microfluidic technology. The unit cell comprises double layer nested ring-split rings formed as microfluidic channels in the Polydimethylsiloxane (PDMS) material using soft lithography techniques. In this structure, only the inner ring has a split region and this region is realized by leaving a gap in the channel filled by an injected liquid metal whereas the outer channel is filled by the liquid metal. Transmission phase of the unit cell is adjusted by implementing rotational phase shifting via manipulating the liquid metal in the channels. X-band unit cell prototypes are fabricated on a glass substrate

Reconfigurable Nested Ring-Split Ring Transmitarray Unit Cell Employing the Element Rotation Method by Microfluidics

A continuously tunable, circularly polarized X-band microfluidic transmitarray unit cell employing the element rotation method is designed and fabricated. The unit cell comprises a double layer nested ringsplit ring structure realized as microfluidic channels embedded in Polydimethylsiloxane (PDMS) using soft lithography techniques. Conductive regions of the rings are formed by injecting a liquid metal (an alloy of Ga, In, and Sn), whereas the split region is air. Movement of the liquid metal together with the split around the ring provides 360 linear phase shift range in the transmitted field through the unit cell. A circularly polarized unit cell is designed to operate at 8.8 GHz, satisfying the necessary phase shifting conditions provided by the element rotation method. Unit cell prototypes are fabricated and the proposed concept is verified by the measurements using waveguide simulator method, within the frequency range of 8-10 GHz. The agreement between the simulation and measurement results is satisfactory, illustrating the viability of the approach to be used in reconfigurable antennas and antenna arrays

Microfluidic Reconfigurable Nested Split Ring-Regular Ring Transmitarray Unit Cell

A reconfigurable, circularly polarized transmitarray unit cell consisting of double layer nested split ring-regular ring formed as microfluidic channels inside Polydimethylsiloxane (PDMS) is designed and fabricated. The operation of the unit cell is based on the element rotation method. The liquid metal is injected into the channels and it forms the conductive part in the structures whereas the split region in the outer ring is air. The movement of the liquid metal together with the split around the outer ring realizes the rotation of the element around its surface normal. The rotation of the element provides a phase shift proportional to the rotation angle and the phase of the transmitted wave can be tuned by controlling this rotation using micropumps. The unit cell is designed to operate at 8.15 GHz and it can be easily scaled to other frequencies