Mostly spread in developing countries Fecal oral transmission

*Mostly spread in developing countries *Fecal – oral transmission from: * Contaminated water * Raw vegetables * Soil * Person to person *Protozoa / Elminths

*Single cell *Trophozoites: living organisms, replication *Cysts: resistant to environment, infectious form *Life cycle can involve different hosts *Example: * Entamoeba hystolytica * Giardia * Cryptosporidium

*Multicellular *Larvae / Adult elminths *Eggs: resistant to environment, infectious form *Example: * Ascaris * Taenia * Hookworm * Strongyloides

WHO Bench Aids https: //www. cdc. gov/dpdx/index. html

*Direct slide microscopic examination (low sensitivity) *Microscopic examination after concentration * Recommended by WHO “Formalin Ether Technique (Ritchie)” *Immunoassays * Microtiter EIA * Rapid tests * Immunofluorescence assays *PCR

* Collection of fecal specimen * Fixation (most commonly Formalin based fixatives) * Disinfection * Morphology preservation * Filtration * Cotton gauze or 0, 25 mm * Food debris elimination * Solvent addition (Ethyl Acetate) * Dissolve fat for easy filtration * Concentration by centrifugation (4 layers) * Sediment microscopic examination * Staining (iodine) * Morphological identification

Tube 13 mm diameter 4 ml of fixative Tube 30 mm diameter 10 ml of fixative

*Optional Item for closing the centrifuge tubes to avoid spills and smell *The cap is perforated so that a Pasteur pipette can be introduced to collect the sediment without the need or removing it *MINI-SYSTEM CAP – 25 RPP 01 500 units

*Optional Screw Caps for closing the centrifuge tubes for long term storage of positive specimens *MINI-SYSTEM SCREW CAPS *25 RPP 04 AB - 500 units

*ECOSAF * Sodium Acetate Formalin fixative * Low formaldehyde content: 0, 58% w/v. *PARAGREEN * Proprietary formulation: does not contain Formaldehyde, Alcohol or Heavy metals

Ø WORK SAFETY Ø IARC classifies formaldehyde as a known human carcinogen Ø Skin sensitizing agent (produces allergies) Ø Vapors are poisonous and heavier than air Exhaust systems are required Ø The human nose can detect concentration of 0, 8 -1 ppm. The US OSHA TWA standards define a 1 ppm exposure limit. (critical concentrations in the air are hardly detected by human smell receptors) Ø Vapors in air can produce explosive mixtures. Ø Vapors have a strong irritant effect on the eyes, lungs and mucosa. Ø Contact with the skin cause sensitization. Ø Swallowing can produce serious irreversible damage and death. Protective measures are required: Respiratory protection, Eye protection, protective clothing and gloves, exhaust systems, medical surveillance of employees. Ø LOGISTIC Ø Danger of fire , High risk of explosion in mixtures between 7% and 73% by volume in air. Ø Classification - Dangerous Goods Ø Formaldehyde oxidizes to produce formic acid which is a highly corrosive substance. Ø ENVIRONMENT Ø Controlled disposal necessary

*Meridian Bioscience * Different closed systems, need for Ethyl Acetate * Ecofix without Formalin, but dangerous *Parasep SF (Solvent Free) - Apacor * Two closed formats, do not require Ethyl Acetate * Saf. EFix fixative, without Formalin, can produce skin allergy

* Contains: Ethanol, Methanol, Zinc Sulfate (heavy metal), Acetic Acid

Parasitology Department Pediatric Hospital Rome Evaluation of concentration techniques of fresh fecal specimens for the quantitative recovery and differentiation of intestinal parasites * Comparison between Mini-System Ecosaf and their system in use: Parasep Formalin (Apacor) * 200 routine specimens and 3 External Quality Control specimens (Neqas) * 170 negative results - 100% agreement * 30 positive results – 97% agreement

Parasitology Department Pediatric Hospital Rome * The Mini-System Ecosaf was found to be easy to use, providing a very clean sediment. * “the sediment concentrated with the Mini-System kit resulted well clarified and therefore particularly suitable for detecting the parasites. From a technical point of view the Mini-System kit resulted particularly valid for its easiness and speed of use. ”

Evaluation of PARAGREEN fixative (Biolife) on different diffusion stages of endoparasites * This study was performed at the Parasitology Laboratory of the Veterinary Faculty of Milan University, to prove the efficacy of this safe fixative. The fact that animal specimens were used is not a limitation, as the parasites are the same which are found in humans. * The study is confirming that the safe Paragreen is working very well just like formalin. Several different types of parasites were included. * “The morphology was well preserved also after 7 days of permanence in the fixative. In summary, the conservation of feces of domestic carnivores in the fixative of Mini-System Paragreen did not produce alterations of various spreading forms of endoparasites (nematodes and protozoa) and those forms maintained all morphological characteristics which allow their correct identification at the optical microscope examination. ”

Mini-System compared to Other company and Home procedure

*Specimens fixed with Formalin and Alcohol based fixatives cannot be further used for other diagnostic tests because of interferences *PARAGREEN fixed specimens were tested by: * Rapid tests Lateral flow assays * Immunofluorescence assays *No interference was found. All samples, both positive and negative provided correct results

*1. A unique disposable system for parasitology concentration * Closed system * No need for Ethyl Acetate *2. A new Formalin free fixative * Same fixation properties as Formalin based fixative * Not dangerous * Minimal workplace safety requirements * No need for exhaust hoods * No special restrictions for transportation and disposal *1+2 = Unique features useful for tenders: * “Closed system with safe fixative for parasite concentration, no Ethyl Acetate is needed”