Comparison of Agar Dilution,Broth Dilution,Cylinder Plate and Disk Diffusion Methods for Evaluation of Anti-leishmanial Drugs on Leishmania promastigotes

Background

Leishmaniases are a group of diseases caused by Leishmania parasites. Growing of drug unresponsiveness in leishmaniasis patients necessitates the development of new drugs and accordingly a suitable assay is needed for evaluation of any modalities. The aim of this study was to compare four drug assays methods, agar dilution, broth dilution, cylinder plate and disk diffusion, for evaluation of anti-leishmanial drugs on Leishmania promastigotes, using glucantime as a currently available drug for treatment of leishmaniasis.

Methods

For broth dilution method, different concentration of glucantime was added to the parasite culture (promastigotes of Leishmania), while in cylinder plate method wells were punched in agar gel and filled with different concentration of drug and zone of inhibition was measured in each well. In disk diffusion method, the parasites were cultivated on the surface of agar; filter paper disks were enriched with various concentration of glucantime and were placed on the surface of agar. In agar dilution method, various concentrations of drug were incorporated onto blood agar and the parasites were cultivated on the surface of the agar.

Results

A direct correlation was found between the drug concentration and size of inhibitory zones in cylinder plate and disk diffusion methods. These two drug assays methods provided much better performance in comparison with broth and agar dilution methods.

Conclusion

Cylinder plate and disk diffusion methods seem to be acceptable methods for susceptibility testing of anti-leishmanial compounds on Leishmania promastigotes.     

Hematologic and Clinical Aspects of Experimental Ovine Anaplasmosis Caused by Anaplasma ovis in Iran

Background

Anaplasma ovis infections can cause clinical symptoms in acute phase and lead to huge economic losses in flocks. The aim of the present study was to investigate the hematological and parasitological changes in experimental anaplasmosis in sheep with Iranian strain of A. ovis.

Method

Five male sheep without any blood parasite infection were selected. One hundred ml heparinized blood was collected from splenectomised sheep that showed 6% A. ovis parasitemia. Inoculums of 20 ml blood were administered intravenously to each test animal. Hematological, parasitological and clinical changes of experimental anaplasmosis were studied in 0-38 days post infection.

Result

Parasitemia was detected 3 days post infection and reached its maximum level on the day 12 of experiment in test animals. Then the parasitemia was declined, but the organism could be found persistently until the last day of study. The red cell counts, packed cell volume and hemoglobin concentration were decreased and mean corpuscular volume was increased significantly during the infection period. Reticulocytosis and basophilic stippling were also detected. No significant changes were observed in total and differential leukocyte count and animal body temperature.

Conclusion

Experimental A. ovis infection in sheep resulted in marked normocytic normochromic anemia at the beginning of the infection which became macrocytic normochromic by the development of the disease. There were negative correlations between parasitemia and RBC, PCV and Hb values, therefore hematological assessment can be considered as a practical diagnostic tool in ovine anaplasmosis.     

Frequency doubling technology and standard automated perimetry in detection of glaucoma among glaucoma suspects

Background

Frequency Doubling perimetry (FDT) has been found to precede visual loss detected by standard automated perimetry (SAP) by as much as four years and the initial development of glaucomatous visual field loss as measured by SAP was found to occur in regions that had previously demonstrated abnormalities on FDT testing.

Methods

A study on 55 glaucoma suspects (determined as per American Academy Guidelines, Preferred Practice Pattern, Oct 2010), was compared to 50 healthy participants (HP). Both glaucoma suspects and HP underwent SAP and FDT in random order. Only reliable fields were compared.

Results

Mean deviation of FDT Matrix was significantly lower than SAP SITA in suspect and healthy group ; two devices showed significant correlation amongst both groups (suspects p = 0.002, healthy p = 0.011). Significant difference was found in PSD of SAP SITA and FDT Matrix (p = 0.001) in the glaucoma suspect group, PSD of FDT Matrix was significantly higher than PSD of SAP SITA in the healthy group (p < 0.001). PSD of SAP SITA significantly correlated with FDT Matrix PSD in glaucoma group (r = 0.579; p = 0.001) but no significant correlation found in healthy group (r = 0.153; p = 0.290). Percentages of normal test locations significantly higher in FDT Matrix compared to SAP SITA in glaucoma suspects and healthy participants.

Conclusion

FDT correlates well with SAP and may be used for patients who are unable to perform well and reliably with SAP but does not show any features of earlier glaucoma changes in this study.     

A study of Hepatitis A and E virus seropositivity profile amongst young healthy adults in India

Background

Various Serosurveys and studies provide ample evidence of differing perspectives regarding epidemiology of HAV and HEV in India. This study was conducted to assess the seroprevalence of HAV and HEV and its associated factors with an aim to provide inputs to planners regarding requirement of HAV vaccine.

Methods

A multi-centric cross sectional survey amongst 4175 healthy trainees (young adults) was carried out in training centres, selected by multistage random sampling, giving equal representation to all regions of India. Sample size was calculated by taking prevalence of HAV seropositivity amongst adults as 60% and alpha 0.05.

Results

Seroprevalence for HAV and HEV was 92.68% (95% CI 91.82, 93.47) and 17.05% (15.90, 18.26), respectively. Logistic regression showed that hand washing without soap, regular close contact with domestic animals, consumption of unpasteurized milk and regular consumption of food outside home were risk factors for HAV (p < 0.05). For HEV, irregular hand washing, consumption of unpasteurized milk and irregular consumption of freshly prepared food were risk factors (p < 0.05).

Conclusion

High level of immunity against HAV among the healthy young adults clearly demonstrates that vaccination against HAV is not required at present in our country. The large proportion being susceptible to HEV points towards the requirement of preventive strategies in the form of safe drinking water supply, hygiene, sanitation, increasing awareness and behaviour change with respect to personal hygiene especially hand and food hygiene.     

Head Injuries

A combination of multiple injury types are typically involved in combat-related head injuries. Innovations in firearms, has led to new types of brain injuries from which we are able to learn much about how the brain responds to trauma. Traumatic brain injury is physical injury to brain tissue that temporarily or permanently impairs brain function. Initial treatment consists of ensuring a reliable airway and maintaining adequate ventilation, oxygenation, and blood pressure. Neurosurgical damage control includes early intracranial pressure control; cerebral blood flow preservation; and prevention of secondary cerebral injury from hypoxia, hypotension, and hyperthemia. Evacuation to the nearest neurosurgeon, avoiding diagnostic delays, and initiating cerebral resuscitation allow for the best chance for ultimate functional recovery.Key Words: Traumatic brain injury, Intracranial pressure, Combat head injury, Primary brain injury, Secondary brain injury     

Spinal Cord Injuries

Spinal Cord injury (SCI) is one of the most devastating and demoralizing ailment for both the patient and the medical practitioner. However, with the better understanding of the pathophysiology, better imaging modalities and emphasis on immobilization and rehabilitation has provided a ray of hope to such patients. The initial care aims at immobilization and evacuation by the classical log roll method and focuses on life-saving procedures. Basic imaging should be augmented with an MRI in doubtful cases. Immobilization either external or internal should be followed by early efforts for rehabilitation. The use of steroids during the acute phase has become controversial. The focus of latest studies has shifted to neuroprotective and regenerative agents.Key Words: Spinal cord injury, Neurological assessment, Magnetic resonance imaging, Methylprednisolone     

Spinal cord injuries

Spinal Cord injury (SCI) is one of the most devastating and demoralizing ailment for both the patient and the medical practitioner. However, with the better understanding of the pathophysiology, better imaging modalities and emphasis on immobilization and rehabilitation has provided a ray of hope to such patients. The initial care aims at immobilization and evacuation by the classical log roll method and focuses on life-saving procedures. Basic imaging should be augmented with an MRI in doubtful cases. Immobilization either external or internal should be followed by early efforts for rehabilitation. The use of steroids during the acute phase has become controversial. The focus of latest studies has shifted to neuroprotective and regenerative agents.     

First report of Taenia acinonyxi (Ortlopp, 1938) in Acinonyx jubatus venaticus from Iran

The Asian cheetah is known as Iranian panther. A four years old female cheetah was killed in a road accident by a truck in Abbas Abad (Biarjamand) County around Shahrood City in Semnan Province, central part of Iran. Two days after the accident the carcass of animal was autopsied and only five cestodes were obtained from its intestine. In inspection of other organs no other helminth was observed. Cestod samples were fixed and stained by carmine acid. Characterization of the cestodes using morphological standard key, identified the cestodes as Taenia acinonyxi.