Forensic analysis of raw meat adulteration using mtDNA

Meat species identification has become essential with the increasing events of frauds like the illegal slaughter of cows, meat adulteration, and substitution. Food scam directly influences public well-being, trade, and wildlife. In Pakistan, donkey meat is used as adulterants for cow meat and is considered Haram in Islamic concepts. In this study PCR, based detection methods are used for identification purposes. The mitochondrial gene cytochrome b has been used in this study to identify the origin of meat specie. Specie-specific primers of cyt b of cow and donkey were used for identification. DNA from different binary ratios of cow and donkey meat was extracted by the phenol-chloroform method. Ratios were made from 1-10 and extracted DNA was subjected to PCR to amplify the target fragment of the cyt b gene. Primers were sensitive to identifying species origin in all meat ratios. Multiplex PCR was designed to identify both species and the results were analyzed by gel electrophoresis. Fragment size of 309bp for cow and 475bp for donkey was observed.Results of the current study conclude that PCR assays, including multiplex PCR, is efficient and has a high sensitivity for even small amount of meat. It is concluded that multiplex PCR is useful and reliable for adulterated meat detection

Pseudosperma quercinum Naseer & Jabeen & Ashfaq & Akbar & Hussain & Khalid 2023, sp. nov.

<i>Pseudosperma quercinum</i> Naseer & Jabeen, <i>sp. nov</i>. (Figure 3 & 4) <p>MycoBank no.: MB 849661</p> <p>Etymology:—The specific epithet “quercinum ” refers to its habitat in oak forests.</p> <p> Holotype:— PAKISTAN. KHYBER PAKHTUNKHWA province, Kurram district, Parachinar, 22 August 2019, <i>Arooj Naseer</i>, (holotype PC-93; LAH35232; GenBank MZ314058 for ITS; MZ314078 for LSU).</p> <p> Diagnosis:—Differs from <i>Pseudosperma umbrinellum</i> by the campanulate to conico-convex pileus with dark brown umbo and yellow-brown fibrillose streaks on creamy white context, a cylindrical stipe with fibrillose, furfuraceous, or almost flocculose apex, ellipsoid to phaseoliform (11.9 × 6.3 µm) basidiospores, and clavate to cylindrical cheilocystidia.</p> <p> Description:— <i>Basidiomata</i> medium-sized. <i>Pileus</i> 3–3.8 cm wide, campanulate to sub-conical, later conico-convex, umbonate, dark brown (7.5R 3/2) at umbo, with yellow-brown (7.5YR 4/6) fibrillose streaks on creamy white context (10Y8/2), dark brown (7.5R 3/2) patches along margins, surface fibrillose, margin striate, creamy white (10Y8/2). <i>Lamellae</i> crowded, adnexed to sinuate, first creamy white (10Y8/2), light silvery brown (10YR 7/2) when mature, edges fimbriate. <i>Lamellulae</i> alternating with lamellae. <i>Stipe</i> 4.3–5.5 × 0.6–0.8 cm, white (5B 9/2) with brown (5Y 5/4) and dark brown tinge at center, cylindrical, central, fibrillose, apex fibrillose, furfuraceous, or almost flocculose, surface dry, white flocculose, hollow, squamulose, base non bulbous and curved. <i>Odor</i> mild, sweet, like honey.</p> <p> <i>Basidiospores</i> [100/3/3] <b>(</b> 8.8–)8.9–13.8(–14.5) × (5.1–)5.6–6.9(–7.1) µm, avl × avw = 11.9 × 6.3 µm, Q = 1.5–2.1, Qav = 1.9, ellipsoid, phaseoliform, guttulate, apiculus small and indistinctive, pale brown to olive greenish in 5 % KOH, thick-walled. <i>Basidia</i> 18.4–32.3 × 8.9–12.7 µm, avl × avw = 26.9 × 10.9 µm, clavate, hyaline in 5 % KOH, thin-walled, 4-sterigmata. <i>Cheilocystidia</i> 18.8–41.6 × 9.8–25.7 µm, avl × avw = 32.2 × 16.2 µm, numerous, narrowly clavate, hyaline in 5 % KOH. <i>Pleurocystidia</i> absent. <i>Pileipellis</i> a cutis of cylindrical hyphae, 5.4–15.8 µm diam., avw = 9.8 µm, septate, branched, hyaline in 5 % KOH. <i>Stipitipellis</i> a cutis of cylindrical hyphae, 3.4–9.9 µm diam., avw = 6.9 µm, septate, branched, hyaline in 5 % KOH. <i>Clamp connections</i> present in all tissues, abundant.</p> <p> Additional materials examined:— PAKISTAN. Khyber Pakhtunkhwa, Swat district, Malam Jabba valley, 1950 m asl., under <i>Quercus incana</i> Roxb. [= <i>Q. oblongata</i> D. Don] in mixed coniferous forest, 10 August 2016, <i>Sana Jabeen MJ1630</i> (LAH35233!; GenBank MZ314059 for ITS; MZ314079 for LSU. Kurram district, Parachinar, 31 July 2018, <i>Arooj Naseer</i>, <i>PC61</i>; (LAH37418!; GenBank OP303370 for ITS).</p> <p> Habitat and distribution:—Solitary on rich loamy soil under <i>Quercus spp</i>. in Kurrum district and Swat district, Khyber Pakhtunkhwa, Pakistan.</p>Published as part of <i>Naseer, Arooj, Jabeen, Sana, Ashfaq, Ammara, Akbar, Mahrukh, Hussain, Syed Iftikhar & Khalid, Abdul Nasir, 2023, Pseudosperma quercinum sp. nov. (Inocybaceae) from the Himalayan forests of Pakistan, pp. 260-270 in Phytotaxa 622 (4)</i> on pages 262-267, DOI: 10.11646/phytotaxa.622.4.3, <a href="http://zenodo.org/record/10084792">http://zenodo.org/record/10084792</a&gt

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