Performance of CO1 and ITS2 nested PCR in molecular identification of ordinary scabies (Sarcoptes scabiei var. hominis)

Scabies is a global disease with a high prevalence, causing morbidity and even mortality, especially in poor and developing countries. However, it is often misdiagnosed due to varied and unspecified lesions. The gold standard technique for diagnosis is a microscopic examination, which requires experienced experts in finding mites, mainly in ordinary scabies. CO1 and ITS2 genes have been widely used in molecular identification to detect Sarcoptes scabiei and its variants. This study aimed to determine and compare the sensitivity and specificity of CO1 and ITS2 S. scabiei genes to the microscopic examination of scabies skin scrapings. The skin scrapings of 52 subjects with scabies diagnosed by anamnesis, physical examination, and dermoscopic examination were examined under a microscope and analyzed by nested PCR. The diagnostic test result showed that the sensitivity of nested PCR of both CO1 and ITS2 genes to micro‐ scope examination was 100%. However, the specificity of both CO1 and ITS2 nested PCR was poor (24% and 0%). Hence, CO1 and ITS2 nested PCR could be more suitable for screening ordinary scabies in humans than the microscopic examination.

MORPHOLOGY AND HISTOLOGY OF THE EURASIAN LYNX (Lynx lynx) PLANUM NASALE

This study reveals the macroscopic and microscopic structures of the Eurasian lynx planum nasale using materials from three dead females obtained from the Sivas Forestry Branch of Agriculture and Forestry Ministry of the Republic of Turkey. To accomplish the purpose, planum nasale was investigated using macroscopic, histological, and scanning electron microscopy (SEM) techniques. The microscopic examination showed that the planum nasale consists of hairless, moist, glabrous skin and resembles a ship anchor with arm, palm, stock, and sickle parts. The planum nasale’s surface is formed by epidermal plates or epidermal ridges, which were separated from each other by primary and secondary fissures showed in SEM and macroscopic figures. Based on the microscopic examination, the Mercel’s cells and nerve ends are located in the basal sheet of the planum nasale’s epidermal layers. In addition, the pores situated on the surface of the epidermal ridges and the dense connective bundles were settled in the dermal layers, based on the SEM examination.Key words: Eurasian lynx (Lynx rufus); morphology; nasal plane; planum nasaleMORFOLOGIJA IN HISTOLOGIJA SMRČKA EVRAZIJSKEGA RISA (Lynx lynx)Izvleček: V študiji so opisane makroskopske in mikroskopske strukture smrčka evrazijskega risa, ki je bila opravljena s proučevanjem tkiv treh mrtvih samic, ki so jih pridobili s pomočjo gozdarske podružnice Sivas Ministrstva za kmetijstvo in gozdarstvo Republike Turčije. Strukturo smrčka so raziskovali z uporabo makroskopskih, histoloških metod ter uporabe vrstičnega elektronskega mikroskopa (SEM). Mikroskopska preiskava je pokazala, da smrček sestavlja brezdlaka, vlažna, gola koža, ki po obliki spominja na ladijsko sidro. Površinski del smrčka tvorijo epidermalne plošče ali grebeni, ki jih ločujejo primarne in sekundarne razpoke, vidne na makroskopskih slikah in s pomočjo vrstične mikroskopije. Na histoloških preparatih so bile v bazalni plasti smrčka epidermisa opazne Merkelove celice in živčni končiči. S pomočjo metode SEM so v plasti epidermisa pokazali pore, ki se nahajajo na površini epidermalnih grebenov in snope togega fibrilarnega veziva, ki segajo v plast dermisa.Ključne besede: Evrazijski ris (Lynx rufus); morfologija; nosna ravnina; smrček

Pharmacognostic and Phytochemical Screening of Datura stramonium by TLC and GC-MS: A Forensic Approach

Forensic Botany helps in linking the location, type of crime, revealing modus Operandi, culprit as well as victim by various circumstantial facts. In the study, Datura stramonium was considered as major plant of Bundelkhand region encountered in poisoning cases in criminal Investigations. Datura plant was identified and its fresh leaves were washed to remove dirt, stored at 40C and were subjected to microscopic examination for botanical features, preliminary screening for alkaloids identification and chromatographic techniques such as TLC and GCMS for qualitative analysis of toxins present in the plant. The result showed that all techniques applied are suitable to identify the plant through botanical derived evidences such as leaf traces, resins, or plant extracts during the crime scene investigations. GCMS technique identified the main toxins and resulted that it contains high concentration of hyoscine and very fatal when administered eventually causes death.

First Report of Powdery Mildew Caused by Podosphaera xanthii on Cucurbita argyrosperma in Mexico

Cucurbita argyrosperma, commonly named as winter or cushaw squash, is highly sought for its seeds, which have important uses in culinary arts. During the autumn 2021, powdery mildew-like signs and symptoms were observed on cushaw squash in several commercial fields located in Cocula, Guerrero, Mexico. Signs were initially appeared as whitish powdery patches on both sides of leaves and then covering entire leaves and causing premature senescence. The disease incidence was estimated to be 80% in about 1000 plants in two fields. The mycelium was amphigenous, persistent, white in color, and occurred in dense patches. A voucher specimen was deposited in the Herbarium of the Colegio Superior Agropecuario del Estado de Guerrero under the accession number CSAEG22. For the morphological characterization by light microscopy, fungal structures were mounted in a drop of lactic acid on a glass slide. Microscopic examination showed nipple-shaped hyphal appressoria. Conidiophores (n = 30) were straight, 100 to 190 × 10 to 12 μm and produced 2 to 6 conidia in chains. Foot-cells were cylindrical, 41 to 78 μm long, followed by 1 to 2 shorter cells. Conidia (n = 100) were ellipsoid-ovoid to barrel-shaped, 29.5 to 39.1 × 19.4 to 22.7 μm, and contained conspicuous fibrosin bodies. Germ tubes were produced from a lateral position on conidia. Chasmothecia were not observed during the growing season. The morphological characters were consistent with those of the anamorphic state of Podosphaera xanthii (Braun and Cook 2012). For further confirmation, total DNA was extracted from conidia and mycelia following the CTAB method (Doyle and Doyle 1990), and the internal transcribed spacer (ITS) region and part of the 28S gene were amplified by PCR, and sequenced. The ITS region of rDNA was amplified using the primers ITS5/ITS4 (White et al. 1990). For amplification of the 28S rRNA partial gene, a nested PCR was performed using the primer sets PM3 (Takamatsu and Kano 2001)/TW14 (Mori et al. 2000) and NL1/TW14 (Mori et al. 2000) for the first and second reactions, respectively. Phylogenetic analyses using the Maximum Likelihood method, including ITS and 28S sequences of isolates of Podosphaera spp. were performed and confirmed the results obtained in the morphological analysis. The isolate CSAEG22 grouped in a clade with isolates of Podosphaera xanthii. The ITS and 28S sequences were deposited in GenBank under accession numbers OL423329 and OL423343, respectively. Pathogenicity was confirmed by gently dusting conidia from infected leaves onto ten leaves of healthy C. argyrosperma plants. Five non-inoculated leaves served as controls. The plants were maintained in a greenhouse at 25 to 35 ºC, and relative humidity of 60 to 70%. All inoculated leaves developed similar signs to the original observation after 10 days, whereas control leaves remained symptomless. Microscopic examination of the fungus on inoculated leaves showed that it was morphologically identical to that originally observed on diseased plants, fulfilling Koch’s postulates. Podosphaera xanthii has been previously reported on C. maxima, C. moschata, and C. pepo in Mexico (Yañez-Morales et al. 2009; Farr and Rossman 2021). To our knowledge, this is the first report of P. xanthii causing powdery mildew on C. argyrosperma in Mexico. This pathogen is a serious threat to C. argyrosperma production in Mexico and disease management strategies should be developed.

The possible alleviating effect of garlic supplement on the neural retina in a rat model of hypercholesterolemia: a histological and immunohistochemical study

The purpose of this work was to prove that oxidative stress is the main mechanism responsible for retinal neurodegenerative changes, subsequent apoptosis, and inflammatory cytokine release in rats fed with a high cholesterol diet (HCD) and determine the role of garlic in alleviating these changes. Forty rats were equally divided into four groups: control, garlic-treated (positive control), HCD, and HCD + garlic-treated (HCD + G). By the end of the experiment (24 weeks) blood samples were collected for assessment of serum lipid profile, oxidative stress parameters, and plasma levels of IL-6 and TNF-α. Both eyes of the rats were enucleated; one was used for light microscopic examination and the other for electron microscopic examination. There was a significant increase in the levels of serum lipids, oxidative stress parameters, IL-6 and TNF-α, and area of expression of caspase-3 in the HCD group compared to both the control and HCD + G groups. Histological examination revealed degenerative changes in all layers of the neural retina in the HCD group. Garlic administration resulted in a significant improvement in the biochemical, immunohistochemical, and histological characteristics of hypercholesterolemic rats. These findings support the hypotheses that garlic has strong antioxidant, anti-apoptotic, and anti-inflammatory properties. Garlic ameliorates the neurodegenerative changes in the neural retina of hypercholesteremic rats.

Epidemiological investigation for brucellosis in dogs of Thrissur

India is endemic to bovine brucellosis, and there is a high potential for transmission of disease from ruminants to dogs. A total of 18 bitches belonging to five different breeds at different stage of abortion (30 days to 65 days of gestation) were selected for this study. Majority of them were showing abortion (88.89 per cent) at 45 to 65 days of the gestation. Microscopic examination of Stamp stained smear obtained from the aborted foetal stomach contents revealed red coccobacillary organisms suggestive of Brucella spp.in three cases. RBPT on paired sera samples on day of presentation and three weeks after abortion showed agglutination within four minutes in five out of 18 female dogs. DNA extracted from the aborted tissues of a RBPT positive Labrador dog yielded amplicons of 193 base pair specific for Brucella spp. on PCR. The results obtained from this study stress the need for screening dogs for canine brucellosis in the current brucellosis surveillance and control programmes.

Uji Diagnostik Filariasis Menggunakan Rapid Diagnostic Test (RDT) Brugia malayi terhadap Pemeriksaan Mikroskopis di Desa Buntoi Kabupaten Gunung Mas, Provinsi Kalimantan Tengah

Gunung Mas Regency, Central Kalimantan Province is one of the endemic filariasis areas with Microfilaria rate of 3.4%. One of the efforts made to control this problem is Mass Drug Administration once a year for 5 years. Currently, the Rapid Diagnostic Test (RDT) method is being developed, a quick and easy diagnostic technique to detect the presence of parasites in the patient's body. This study aims to determine the results of the filariasis diagnostic test using the Brugia malayi RDT on the microscopic examination in Buntoi Village, Gunung Mas Regency, Central Kalimantan Province. This research is a descriptive study with a cross-sectional approach with the research subjects all residents of Buntoi Village with inclusion and exclusion criteria totaling 161 samples. Collecting data was carried out by examination and interviews with questionnaires. Data analysis by calculating the microfilaria rate, sensitivity and specificity and calculating the frequency distribution of research variables. Data is presented in percentage form and displayed in tabular form. The results of the diagnostic study of B. malayi RDT and the microscopic examination were the same, i.e all were negative and no microfilariae were found. The diagnostic test for filariasis RDT Brugia malayi on microscopic examination (SDJ) obtained 0% sensitivity, 100% specificity, 0% Positive Predictive Value and 100% Negative Predictive Value. The level of public knowledge about filariasis includes 61% good category, knowledge of MDA 40% good category and knowledge about prevention of filariasis in good category 53%.

The Anthelmintic Activity Of Etanol Extract of Porang Tubers Against Fasciola gigantica In Vitro

This study aims to determine the anthelmintic activity ethanol extract of porang tubers (A. oncophillus) against F. gigantica worms in vitro. There were five treatments and each treatment was done in five replications and used 10 F. gigantica. The treatments of this research were K- with CMC Na 1%, K+ with Albendazole 2.4 mg/ml, P1 with extract concentration 5%, P2 with extract concentration 10%, P3 with extract concentration 20%. The results showed that the extract of Porang tuber (A. oncophillus) had an anthelmintic effect against F. gigantica worms in vitro. In the extract with a concentration of 20%, there were anthelmintic properties that almost the same as Albendazole. The higher the extract concentration, the higher the anthelmintic properties. The longer the immersion time, the higher the number of dead worms. The morphological changes was evaluated by light microscopic examination and the results showed many histopathological changes on the morphology of F. gigantica. The results indicate that the possible use of the tubers as a potential anthelmintic against F. gigantica.

Pathological Features of COVID-19 Pneumonia Diagnosed Following an Autopsy

After initial emergence at the end of 2019, coronavirus disease 19 (COVID-19) rapidly spread to become a pandemic. The causative agent is recognized as a novel coronavirus, named as severe acute respiratory syndrome coronavirus 2, which is known to primarily affect the respiratory system. We present the case of a 60-year-old man who was diagnosed to have COVID-19 pneumonia following an autopsy. Microscopic examination of the lung tissue showed acute (exudative) phase of diffuse alveolar damage, inflammatory infiltrates in alveolar spaces and interstitium, atypical pneumocytes with prominent nucleoli and expanded cytoplasm, and thrombi in small arterioles. Some megakaryocytes were observed, which were positive for immunohistochemical stain for CD61. The pathological findings observed in this case were consistent with those of previously published reports of COVID-19 pneumonia. This is the first case of COVID-19 pneumonia diagnosed with an autopsy in Korea.

Synergistic Cytotoxic Effect of Honey Bee Venom and Cisplatin on Tongue Squamous Cell Carcinoma Cell Line

BACKGROUND: Tongue cancer is one of the most common head and neck cancers in the world. Nowadays, natural compounds are important resources of many anti-cancer drugs. Venom from honey bees possesses potent anti-cancer activities. Cisplatin is a chemotherapeutic drug that has been used for decades to treat cancer cells. Recently, combination therapy has been a popular treatment choice for cancer patients. AIM: This study was conducted to evaluate the synergistic cytotoxic effect of honey bee venom (BV) and cisplatin on tongue squamous cell carcinoma 25 (SCC-25) cell lines. METHODS: The cytotoxic effect was determined using methyl thiazol tetrazolium assay, microscopic examination, real-time polymerase chain reaction (RT-PCR), and statistical analysis. RESULTS: The findings revealed that the cytotoxic potential of the tested drugs on SCC-25 cells was dose-dependent. Microscopic examination showed that BV and cisplatin alone and in combination mainly produced apoptotic cell death. Regarding RT-PCR results, P53 and caspase-3 expression levels were significantly increased in SCC-25-treated cells (p = 0.0001). CONCLUSION: The combined use of BV and cisplatin induced a marked synergistic cytotoxic effect on SCC-25 cell line.