Microbiological Quality of Selected Local and Imported Non-Sterile Pharmaceutical Products in Dar es Salaam, Tanzania

Figures & data

Table 1 Sampling Details

Product Type/ Dosage Form	Active Ingredient/Generic Name	Product Origin (Local/Imported)	Minimum Quantity (Units) Collected per Batch	Sample Code (Collected Bulk Samples Randomly Picked in the Lab to Make Pooled Test Samples)	Amount Required per Sample for One Test (As per Harmonised Pharmacopeial Methods)
					Total Aerobic Microbial Count (TAMC)	Total Yeast and Mould Count (TYMC)	Tests for Specified Microorganisms
Tablets	Paracetamol	Local	100	1	10g	10g	10g
		Imported	100	2	10g	10g	10g
	Sildenafil citrate	Local	100	3	10g	10g	10g
		Imported	100	4	10g	10g	10g
Capsules	Cough capsules	Local	100	5	10g	10g	10g
		Imported	100	6	10g	10g	10g
Syr/Susp	Paracetamol	Local	6	7	10mls	10mls	10mls
		Imported	6	8	10mls	10mls	10mls
	Cough syrup/suspension	Local	6	9	10mls	10mls	10mls
		Imported	6	10	10mls	10mls	10mls

Table 2 Recommended Acceptance Criteria for Microbiological Quality of Non-Sterile Dosage Forms (European Pharmacopeia)

Route of Administration	Total Aerobic Microbial Count (CFU/g or CFU/ mL)	Total Combined Yeasts/Moulds Count (CFU/g or CFU/ mL)	Specified Microorganism
Non-aqueous preparations for oral use	10^3	10^2	Absence of Escherichia coli (1 g or 1 mL)
Aqueous preparations for oral use	10^2	10^1	Absence of Escherichia coli (1 g or 1 mL)
Rectal use	10^3	10^2	-
Oromucosal use Gingival use Cutaneous use Nasal use Auricular use	10^2	10^1	Absence of Staphylococcus aureus (1 g or 1 mL) Absence of Pseudomonas aeruginosa (1 g or 1 mL

Table 3 Average Total Counts of the Microbial Contaminants Found in Selected Locally Produced and Imported Non-Sterile Pharmaceutical Products

Product Type/ Dosage Form	Active Ingredient/Generic Name	Product Origin (Local/Imported)	Sample Code	Average Total Aerobic Microbial Count (TAMC- CFU/g or CFU/mL)	Average Total Yeast and Mould Count (TYMC- CFU/g or CFU/mL)	Quality Remarks with Respect to TAMC	Quality Remarks with Respect to TYMC
Tablets	Paracetamol	Local	1	6.5 x 10^3	1.5 x 10^2	Fail	Pass
		Imported	2	1.4 x 10^4	5.5 x 10^3	Fail	Fail
	Sildenafil citrate	Local	3	6.0 x 10^2	4.5 x 10^2	Pass	Fail
		Imported	4	2.5 x 10^3	1.5 x 10^2	Fail	Pass
Capsules	Cough capsules	Local	5	3.0 x 10^2	1.5 x 10^2	Pass	Pass
		Imported	6	1.3 x 10^3	5.0 x 10^3	Pass	Fail
Syr/Susp	Paracetamol	Local	7	8.0 x 10^3	2.4 x 10^3	Fail	Fail
		Imported	8	5.5 x 10^3	1.3 x 10^4	Fail	Fail
	Cough syr/susp	Local	9	1.9 x 10^3	1.8 x 10^3	Fail	Fail
		Imported	10	8.5 x 10^3	1.0 x 10^3	Fail	Fail

Table 4 Identification of Bacterial Contaminants Isolated from the Products

Product Type/ Dosage Form	Active Ingredient/Generic Name	Product Origin (Local/Imported)	Sample Code	Colony Description	Gram Staining	Specified Biochemical Test(s)	Conclusion
Tablets	Paracetamol	Local	1	Yellow, slightly flat, NLF^a	GNR^b	Oxidase (+)	Pseudomonas aeruginosa confirmed
		Imported	2	No growth on selective medium	–	–	No pathogenic bacteria
	Sildenafil citrate	Local	3	Yellow, NLF	GNR	Oxidase (+)	Pseudomonas aeruginosa confirmed
				Red, LF^c	GPC^d	Catalase (+), DNase (-), Novobiocin sensitive	Staphylococcus epidermidis confirmed
		Imported	4	Yellow, NLF	GNR	Oxidase (+)	Pseudomonas aeruginosa confirmed
Capsules	Cough capsules	Local	5	Yellow, flat, NLF	GNR	Oxidase (+)	Pseudomonas aeruginosa confirmed
		Imported	6	Pink/red, slightly raised, LF	GPC	Catalase (+), DNase (-), Novobiocin sensitive	Staphylococcus epidermidis confirmed
Syr/Susp	Paracetamol	Local	7	Mucoid yellow on CLED^e, LF	GNR	Oxidase (-), Urease (-), Citrate (+), KIA^f [Gas (+), H2S (-), Reaction(acid/acid)], SIM^g [H2S (-), Indole (-), Motility (-)]	Klebsiella pneumoniae confirmed.
		Imported	8	Pink, LF	GPC	Catalase (+), DNase (-), Novobiocin sensitive	Staphylococcus epidermidis confirmed
	Cough syr/susp	Local	9	LF	GPC	Catalase (+), DNase (-), Novobiocin sensitive	Staphylococcus epidermidis confirmed.
				LLF^h	GPC	Catalase (+), DNase (-), Novobiocin sensitive	Staphylococcus epidermidis confirmed
		Imported	10	Yellow, NLF	GNR	Oxidase (+)	Pseudomonas aeruginosa confirmed

Notes: ^aNon-Lactate Fermenter; ^bGram-Negative Rods; ^cLactate Fermenter; ^dGram-Positive Cocci; ^eCystine Lysine Electrolytes Deficiency; ^gSulphur Indole Motility; ^hLate Lactate Fermenter.

Abbreviation: ^fKIA, Kligler’s Iron Agar.

Table 5 Identification of Fungal Contaminants Isolated from the Products

Product Type/ Dosage Form	Active Ingredient/Generic Name	Product Origin (Local/Imported)	Sample Code	Colony Description (Macroscopic Identification)	Microscopic Features	Suspected Organism
Tablets	Paracetamol	Local	1	Greenish-yellow, white margins. Reverse- brownish	Non-septate hyphae, conidiophores resembling sunflower,	Aspergillus flavus
		Imported	2	Greenish-yellow, white margins. Radical fold radiating from the centre	Clubbed vesicles	Aspergillus fumigatus
	Sildenafil citrate	Local	3	Dark-green, yellow	Spherical vesicles surrounded by double-row spores	Aspergillus flavus
				Brownish-yellow	Clubbed vesicles	Aspergillus fumigatus
		Imported	4	Greenish-yellow	Spherical vesicles surrounded by double-row spores	Aspergillus flavus
Capsules	Cough capsules	Local	5	Dark-brown, greenish-yellow	Spherical vesicles surrounded by double-row spores	Aspergillus flavus
		Imported	6	Dark-brown, yellow	Septate with clear round vesicles	Aspergillus flavus
				Greenish-yellow	Clubbed vesicles	Aspergillus fumigatus
Syr/Susp	Paracetamol	Local	7	Greenish with radial fold	Conidiophores forming branches- brush-like	Penicillium spp
		Imported	8	Greenish with radial fold	Conidiophores forming branches- brush-like	Penicillium spp
	Cough syr/susp	Local	9	Dark-green, yellow with radial folds	Round vesicles with spores	Aspergillus flavus
		Imported	10	Greenish-yellow	Non-septate mycelium with round vesicles resembling sunflower	Aspergillus flavus

Table 6 Susceptibility Patterns (%) of the Isolated Pathogens Against Common Cell-Wall-Targeting Antibiotics and Protein Synthesis Inhibitors

Isolate		P. aeruginosa			S. epidermidis			K. pneumonia
Susceptibility		S^a	I^b	R^c	S	I	R	S	I	R
Cell-wall-targeting antibiotics	AMX^d	0	0	100	0	0	100	0	0	100
	AMC^e	0	20	80	20	40	40	0	0	100
	AMP^f	20	0	80	20	20	60	0	0	100
	PIP^g	20	60	20	60	0	40	0	0	100
Penicillins overall		10	20	70	25	15	60	0	0	100
	CRO^h	100	0	0	60	20	20	0	0	100
	CAZ^i	60	0	40	0	20	80	0	0	100
	FOX^j	20	40	40	100	0	0	0	0	100
Cephems overall		60	13.3	26.7	53.4	13.3	33.3	0	0	100
	VAN^k	40	0	60	100	0	0	0	0	100
Class Overall		32.5	15	52.5	45	12.5	42.5	0	0	100
Protein synthesis inhibitors	GEN^l	60	0	40	60	0	40	0	0	100
	CHL^m	20	20	60	60	0	40	0	0	100
	NIT^n	40	0	60	100	0	0	0	0	100
	ERY^o	0	40	60	20	20	60	0	0	100
	TS^p	0	0	100	20	0	80	0	0	100
	NA^q	0	20	80	0	0	100	0	100	0
Class Overall		20	13.3	66.7	43.3	3.3	53.4	0	16.7	83.3
Overall		27.1	14.3	58.6	44.3	8.6	47.1	0	7.1	92.9

Notes: ^aSensitive; ^bIntermediate; ^cResistant; ^dAmoxicillin; ^eAmoxicillin- Clavulanic acid; ^fAmpicillin; ^gPiperacillin; ^hCeftriaxone; ⁱCeftazidime; ^jCefoxitin; ^kVancomycin; ^lGentamicin; ^mChloramphenicol; ⁿNitrofurantoin; ^oErythromycin; ^pTrimethoprim-Sulfamethoxazole; ^qNalidixic acid.

Abbreviations: AST, antibiotic susceptibility testing; CFU, colony forming unit; EP, European Pharmacopoeia; GMP, good manufacturing practices; MAC, maximum acceptable count; USP, MAL, maximum acceptable level; NSP, non-sterile pharmaceutical(s); QA, quality assurance; QC, quality control; Susp, suspension; Syr, syrup; TAMC, total aerobic microbial count; TBC, total bacterial count; TFC, total fungal count; TVC, total viable count; TYMC, total combined yeast and mould count; United States, Pharmacopoeia.